Objective To evaluate the expression of Toll-like receptor(TLR)on the monocyte- derived dendritic cells(DC)from chronic hepatitis B(CHB)patients and to analyze the expression pro- file and significance of the TLR such as TLR3,TLR4,TLR?,TLR8 and TLRg,which are associat- ed with immune response to viral infection.Methods Peripheral blood mononuclear cell(PBMC) centrifugated by the hydroxyethyl starch(HES)centrifugation were cultured and induced into DC by granulocyte-maerophage colony stimulating factor(GM-CSF)and interleukin-4(IL-4),and their mor- phology and phenotype were detected by the inverted microscope and flow cytometry respectively. Monocyte-derived DC were obtained from 10 chronically hepatitis B virus(HBV)-infected patients and 15 healthy volunteers.TLR3,TLR4,TLR7,TLRS,TLR9 expression on immature and mature DC were analyzed by FACS Calibur.DC was pulsed with HBcAg on day 3 and 5,then DC maturation and ability to process HBcAg and to stimulate autogeneic T cells were evaluated.Results Monocyte- derived DC developed different TLR expression patterns as they went through different maturation stages.TLR7,TLR8 expressions on immature DC and TLR3,TLR7 expressions on mature DC were lower in CHB than in control(for TLR7,TLR8 expression on immature DC:75.9%,1.0%vs 98.4%,15.4%,P

During the process of growth, harvesting, transportation, processing and storage, Chinese herbal medicines (CHMs) can be easily contaminated by fungi and their metabolites like mycotoxins, which not only express negative effects on the quality and safety of CHMs and their processed products, but also pose great threats to human health. Now, some chemical synthetic fungicides have been frequently used to control the growth of fungi and accumulation of mycotoxins in the preservation of CHMs. However, the concentration and type of chemical fungicides allowed for postharvest application are restricted due to the disadvantages of their high residual toxicity, long degradation period and pollution to the environment and so on. Therefore, it is critical to research and develop some highly effective, safe and non-toxic, natural, environment-friendly fungistatic agents from plants to prevent CHMs from being contaminated by fungi and mycotoxins. The paper reviews mycotoxins and their harmfulness, the effective compounds of fungistatic plants as well as the antifungal mechanism to provide scientific evidences for developing novel and effective fungistatic agents plants. Then, the application prospect of fungistatic agents from plants in the preservation of CHMs was discussed.
Animals ; Fungi ; drug effects ; metabolism ; Fungicides, Industrial ; pharmacology ; Humans ; Mycotoxins ; metabolism ; toxicity ; Plant Diseases ; microbiology ; prevention & control ; Plant Extracts ; pharmacology ; Plants, Medicinal ; chemistry ; microbiology ; Preservation, Biological ; methods

OBJECTIVETo assess the in-stent lumen visibility and image quality of coronary stents by dual-source computed tomography (DSCT) coronary angiography, and the diagnostic accuracy of DSCT in the detection of coronary in-stent restenosis.

METHODSDSCT was performed at 147 stents in 78 patients at an interval of (21.8?22.2) months after coronary stent implantation. Axial multi-planar reconstruction of the stents and curved-planar reconstruction through the median of the stents were evaluated for stent image quality on a 5-point scale, and the stent lumen diameters were detected. Thirty out of these 78 patients underwent conventional coronary angiography within one month after CT angiography. The patency of 60 stents were independently evaluated by two blinded readers.

RESULTSImage quality was good to excellent on average score (1.6?0.6) . Stent image quality score was correlated with stent diameter, stent location, and heart rate. All stents were assessable in lumen visibility with an average visible lumen diameter percentage of (72.2?12.2) %. Visible lumen diameter percentage was correlated with stent diameter and stent location. For the stents with calcified plaques, the visible lumen diameter percentage at the calcified site was significantly lower than that at the non-calcified site (P<0.001) . Compared with the conventional coronary angiography, 12 out of 14 in-stent stenoses were correctly detected. The sensitivity, specificity, positive predictive value, and negative predictive value for the detection of in-stent stenosis was 85.7%, 95.7%, 85.7%, and 95.7%, respectively. For stents whose diameter >0.275cm, the sensitivity, specificity, positive predictive value, and negative predictive value were all 100%. The agreement between CT findings and coronary angiography results was 93.3%, and it was correlated with stent diameter and heart rate.

CONCLUSIONSUsing a DSCT, coronary stent lumen is partially visible and the image quality is high. Stent diameter and location can influence the stent lumen visibility and image quality. DSCT has a high diagnostic accuracy for the detection of in-stent restenosis and may be a valuable modality for the follow-up of coronary artery stent patency."

Aged ; Coronary Angiography ; methods ; Coronary Restenosis ; diagnostic imaging ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Sensitivity and Specificity ; Stents ; Tomography, X-Ray Computed ; methods ; Vascular Patency

Objective:To improve the genetic stability of HBV gene in transgenic mice.Methods:HBV transgenic mice were bred by backcross and double cross.The HBV gene expression and replication were studied with real-time PCR,ELISA and chemiluminescence.Results:The HBV transgenic mice have stably bred to 23rd generation.The serum HBsAg level is 4122.31?2044.74IU/ml;The rate of HBV transgenic mice whose serum HBV DNA reach 104~106copies/ml was 93.93%.The HBV replication and expression were improved markedly.There is no difference between male and female mice about serum HBsAg level.Conclusion:After breeding the HBV gene was expressed stably with high-level in transgenic mice.

BACKGROUND: C-telopeptide and N-telopeptide cross-linked collagen type Ⅰ (CTx and NTx, respectively) are specific biochemical bone markers that can reflect bone formation and resorption. OBJECTIVE: To analyze the association of CTx with disuse osteoporosis. METHODS: Male Sprague-Dawley rats, weighing 180-220 g, were randomly divided into control and disuse osteoporosis groups. Right hind limbs of the rats in the disuse osteoporosis group were immobilitzed for 4 weeks by ankle-tail fixation to establish the rat model of disuse osteoporosis. Peritoneal venous blood was collected before and after modeling, and the femur was then removed to measure the serum CTx level and bone mineral density of the bilateral femurs. RESULTS AND CONCLUSION: The serum CTx level did not differ significantly between groups before modeling (P > 0.05). At 4 weeks after modeling, the serum CTx level in the disuse osteoporosis group was significantly higher than that in the control group and at baseline (P <0.01). The serum CTx level showed no significant change in the control group before and after modeling (P > 0.05). The increment of serum CTx in the disuse osteoporosis group exhibited a negative correlation with the bone mineral density of the bilateral femurs (r=0.426, P < 0.01). The bone mineral density of the right femur in the disuse osteoporosis group was significantly lower than that of the left one in the disuse osteoporosis group and the right one in the control group (P < 0.01), and there was no significant difference in the bone mineral density between left and right femurs in the control group (P > 0.05). These results imply that the model of disuse osteoporosis by ankle-tail fixation is established successfully. Disuse osteoporosis can promote the production of CTx further reducing bone mineral density; CTx is positively correlated with the degree of bone loss, so it can be used for therapeutic assessment and diagnosis of osteoporosis.

OBJECTIVETo study the immunoregulatory effect of hepatitis B virus (HBV) e antigen (HBeAg) on peripheral blood monocytes (PBMCs).

METHODSPBMCs were isolated from patients with chronic hepatitis B (CHB; both HBeAg- and HBeAg+) and healthy controls, and cultured with recombinant HBeAg. The HBeAg-induced changes in expression of PD-1/PD-L1 were measured by flow cytometry of the cells and in secreted cytokines were measured by enzyme-linked immunosorbent assay of the supernatants. Comparisons between two groups were made by the independent-samples t-test; the relationship between PD-1/B7-H1 level and HBV DNA copy number was evaluated by Spearman's correlation analysis.

RESULTSExposure to HBeAg led to a significant decrease in CD3+CD4+ T lymphocyte-specific expression of IFNa for both the CHB patients' and healthy controls' samples (t = 2.382 and -4.190 respectively, P less than 0.01). For the HBeAg- CHB patients' and healthy controls' samples, the HBeAg exposure led to increased levels of secreted cytokines IL-6, IL-10 and TNFa (t = 2.504, 3.583 and 4.324, P less than 0.01 and t = 3.542, 6.246 and 5.273, P less than 0.01 respectively) and of CD14+ PBMC-specific expression of PD-L1 (t = 4.815 and 3.454, P less than 0.05 respectively). Compared to the HBeAg-negative CHB patients' and healthy controls' samples, the HBeAg+ CHB patients' samples had significantly lower CD3+CD4+ T cell-specific expression of IFNa (t = -3.177 and -4.541, P less than 0.01 respectively), but significantly higher levels of secreted IL-4 (t = 3.382 and 4.393, P less than 0.01 respectively), of CD3+ T cells-specific expression of PD-1/PD-L1 (t = 4.755, 2.942 and 4.518, 4.595, P less than 0.01 respectively), and of CD14+ T cells-specific expression of PD-L1 (t = 5.092 and 5.473, P less than 0.01 respectively). The CD3+ T cells-specific expression of PD-L1 was significantly higher in the samples from HBeAg- CHB patients than from the healthy controls (t = 3.214, P less than 0.01).

CONCLUSIONHBeAg was able to down-regulate the production of Th1-type cytokines (IFNgamma), and up-regulate the secretion of Th2-type cytokines (IL-6, IL-10) and the expression of PD-1/PD-L1on monocytes. These changes are conducive to the formation of immune tolerance to HBV. Therefore, HBeAg may play an important role in immune tolerance to chronic HBV infection.

Adult ; Case-Control Studies ; Cells, Cultured ; Female ; Hepatitis B e Antigens ; genetics ; immunology ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Interferon-gamma ; immunology ; Interleukin-10 ; immunology ; Interleukin-6 ; immunology ; Leukocytes, Mononuclear ; immunology ; metabolism ; Male ; Middle Aged ; Recombinant Proteins ; immunology ; Th1 Cells ; immunology ; Th1-Th2 Balance ; Th2 Cells ; immunology

Objective To investigate the association of post-stroke depression (PSD) with gene polymorphisms of catechol-O-methyl transferase (COMT) Val1 08/158Met and dopamine transporter 40bp variable number of tandem repeats (VNTR) in dopamine metabolism system. Methods Sixty-eight patients with PSD and 91 patients only suffered from stroke, admitted to our hospital from January 2010to June 2010, were chosen; the gene polymorphisms ofCOMT Val108/158Met and DAT 40 bp VNTR were analyzed by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results The genotypes of COMT gene amplifications were wild type (G/G),homozygous mutant type (A/A) and heterozygous type (A/G); 7 repeated genotypes (7/7, 9/7, 10/7, 10/9,10/10, 11/10 and 11/11) were noted in the DA T gene amplifications; frequencies of COMT alleles and genotypes were significantly different between the 2 groups (x2=5.703, P=0.017;x2=6.489, P=0.039). The frequencies of COMT alleles and genotypes were significantly different between the 2 female groups (x2=4.610, P=0.032;x2=6.547, P=0.024), but no significant differences were found between the 2 male groups (P＞0.05). The frequencies and heterozygosity of DAT alleles and genotypes showed no obvious differences between the 2 groups (P＞0.05). Conclusion The gene polymorphism of COMT Val108/158Met may be associated with PSD, while that of DAT 40bp VNTR is not.

OBJECTIVETo establish and optimize a real time RT-PCR system for determining the transcript levels of CatSper1 in human and mouse mature spermatozoa containing microamount of RNA.

METHODSTotal RNA of human and mouse mature spermatozoa was isolated by using TRIzol reagent and reversely transcribed to complementary DNA respectively. Primers for real time RT-PCR were designed in the homologous area of the human and mouse CatSper1 mRNAs. Human sperm complementary DNA was used as the template to the optimize the conditions for SYBR Green I real time RT-PCR, including annealing temperature, Mg2+ concentration, fluorescence measurement temperature and the ratio between forward and reverse primers. The standard curve was constructed with serial dilutions of complementary DNA from human sperm to ascertain the amplification efficiency of SYBR Green I real time PCR and to quantitate the CatSper1 mRNA levels in the human and mouse mature spermatozoa.

RESULTSThe optimal conditions for real time RT-PCR, that is, annealing temperature, Mg2+ concentration and the ratio between forward and reverse primers were 63 degrees C, 3.0 mmol/L and 1:1 respectively. The fluorescence measurement temperature was 88 degrees C. The standard curves were Y = -3.402 log (X) + 25.99 and Y = -3.409 log(X) + 24.09 in the human sperm cDNA and mouse sperm cDNA as the template, with amplification efficiency of 96.8% and 96.5% respectively. The R2 value (an indicator of the quality of the fit of the standard curve to the standard data points plotted) of both standard curves was 0.998. The CatSper1 mRNA levels in the human and mouse mature spermatozoa could be determined according to the standard curve.

CONCLUSIONThe general RT-PCR system, by adding SYBR Green I and optimizing its conditions, could be used to quantitate the mRNA levels in both human and mouse mature spermatozoa.

Animals ; Calcium Channels ; genetics ; Humans ; Male ; Mice ; Organic Chemicals ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Spermatozoa ; metabolism

OBJECTIVESIn order to investigate the relationship among the toll-like receptor 2 (TLR2), hepatitis B e antigen and HBV DNA, the expression levels of TLR2 on peripheral blood monocytes of chronic hepatitis B (CHB) patients as well as on their monocytes stimulated by ligand of TLR2 (Pam3CSK4) and HBeAg were analyzed.

METHODSSixty-eight adults with CHB were enrolled, including 37 HBeAg-positive patients, 17 HBeAg-negative and HBV DNA negative patients, and 14 HBeAg-negative and HBV DNA positive patients. Sixteen healthy volunteers were also studied as controls. TLR2 expression levels on their peripheral blood monocytes stimulated with Pam3CSK4 or not stimulated were analyzed by FACS Caliber. The relationship of the expression levels of TLR2, HBeAg and HBV DNA were also analyzed. The level of TLR2 on peripheral blood monocytes of healthy volunteers and HBeAg-negative CHB patients stimulated by HBeAg was examined for six hours.

RESULTSThe TLR2 expression levels on CD14+ cells were significantly reduced in HBeAg-positive patients (47.57%+/-21.40 %) compared to both healthy volunteers (76.51%+/-7.46%) and HBeAg-negative patients (HBV DNA positive group 73.2%+/-14.2%, HBV DNA negative group 75.2%+/-11.3%); but there was no difference between those of the HBeAg-negative patients and the healthy volunteers. Expression levels of TLR2 on monocytes stimulated by TLR2 ligand in HBeAg-positive patients were obviously increased, and reached the basic levels of the healthy volunteers and the HBeAg-negative patients. The expression levels of TLR2 on monocytes stimulated by HBeAg of the healthy volunteers and the HBeAg-negative patients were markedly reduced.

CONCLUSIONSIn the presence of HBeAg, HBV down-regulates the expressions of TLR2 on CD14+ cells from peripheral blood, and there is no correlation between HBV-DNA and TLR2. Pam3CSK4 can boost the TLR2 expression in HBeAg-positive patients. The proposed interaction between HBV and TLR2 may provide an important clue to explain the reasons of the establishment of persistent HBV infection.

Case-Control Studies ; DNA, Viral ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; immunology ; metabolism ; Humans ; Lipopolysaccharide Receptors ; metabolism ; Monocytes ; metabolism ; Toll-Like Receptor 2 ; metabolism

OBJECTIVETo evaluate image quality (IQ) and radiation exposure of coronary computed tomographic angiography (CTA) with prospectively electrocardiographic (ECG) triggered high-pitch spiral acquisition using dual source CT.

METHODSTotally 75 consecutive patients with a stable heart rate (HR) ≤65 bpm underwent coronary CTA. patients were divided into two groups according to their HR (group A HR≤60 bpm, group B HR >60 bpm to≤65 bpm) . A dual-source CT scanner was used (0.6mm collimation, 0.28s rotation time, 80~100 kV, 370 mAs/rot) . Data acquisition was prospectively ECG-triggered at 60% of the R-R interval with a pitch of 3.4. Images were reconstructed with 75ms temporal resolution, 0.75mm slice thickness and 0.5mm increment. IQ was evaluated using a four-point scale (1=excellent, 4=unevaluable) .

RESULTSThe mean HR and scan time of all patients was (57.2 ± 4.8) bpm and (0.42 ± 0.02) s. Of 1103 coronary artery segments, 934 (84.7%) had an IQ score of 1, 135 (12.2%) score of 2, 18 (1.6%) score of 3,and 16 (1.5%) were rated as unevaluable. There was no significant difference between the two groups in IQ [mean score (1.19 ± 0.52 vs. 1.22 ± 0.55;Z=-1.107,P=0.268) . The rate of evaluable segments showed no significant difference between the two groups (98.5% vs. 98.6%;X2=0.000,P=1.000) . Mean dose-length product of all patients was (67.2 ± 30.4) mGy × cm, mean effective dose was (0.94 ± 0.43) mSv.

CONCLUSIONIn patients with a stable HR of 65 bpm or less, prospectively ECG-triggered high-pitch spiral CT acquisition provides high IQ at low radiation dose.

Aged ; Bradycardia ; diagnostic imaging ; Coronary Angiography ; methods ; Female ; Humans ; Male ; Middle Aged ; Quality Control ; Radiation Dosage ; Radiographic Image Interpretation, Computer-Assisted ; Tomography, Spiral Computed ; methods