Objective To detect genetic polymorphism of Toll like receptor 2 (TLR2) R753Q and Toll like receptor 4 (TLR4) D299G and T399I ,and to analyze its role and mechanism in the occurrence and development of colorectal carcinoma .Meth‐ods Totally 256 cases of patients with colorectal carcinoma and 256 cases of healthy control individuals were collected .The geno‐types of TLR2 R753Q ,TLR4 D299G and TLR4 T399I were detected by using PCR‐RFLP method .The levels of IL‐1α,IL‐8 ,MIP‐1αand MCP‐1 protein were detected by using ELISA .Results It is shown that TLR4 T399I was associated with the risk of color‐ectal cancer .The frequency of CT combined TT genotype of T399I in case group and control group was 31 .2% and 18 .7% ,respec‐tively .The frequency of T allele in case group and control group was 18 .2% and 10 .2% ,respectively ,and difference was statistical‐ly significant(P<0 .05) .Individuals with T allele of T399I showed a 2 .534‐fold increase in colorectal cancer risk compared with the T399I C allele(95% CI:1 .462-2 .734 ,P<0 .01) .There were significant differences in the expression levels of IL‐1αand MCP‐1 in colorectal carcinoma tissues of different genotypes .The expression level of IL‐1α in patients with CT combined TT genotype of T399I and those with CC genotype was (36 .97 ± 21 .43) and (22 .27 ± 17 .89)pg/mg respectively ,and the expression level of MCP‐1 was (24 .57 ± 17 .74) and (12 .91 ± 9 .78)pg/mg respectively .Conclusion T399I TLR4 is associated with the occurrence and de‐velopment of colorectal carcinoma ,and the risk of colorectal carcinoma in individuals with T genotype is significantly increased ,in which monitoring the expression of IL‐1 and MCP‐1 in colorectal carcinoma tissues might be the mechanism .

Objective To have knowledge of the exact cause of massive osteolysis. Methods Hair of patients from Xinjiang province was collected and 14 trace elements were determined by inductively coupled plasma mass spectrometryICP-MS. Results Trace elements imbalance in the body of patients was disturbed. Chromium and zinc which are benefit to the growth of the bones were only 0.5 ?g/g and 40 ?g/g respectively that were much lower compared with the healthy persons cadmium was much higher than the limit level in healthy person. Moreover the quantity and ratio of potassium and sodium in the patients were obvious abnormal. Conclusion According to the result of the present paper may be the environmental and dietary factors play an important role in pathogenesis of this disease.

Objective To compare the effects of remifentanil infused at different rates on median effective target plasma concentration (EC50) of propofol inhibiting responses to laryngeal mask airway (LMA) insertion and determine the optimum infusion rate of remifentanil when used for fiberoptic bronchoscopy in pediatric patients.Methods Eighty-four ASA Ⅰ or Ⅱ pediatric patients,aged 7 months-3 years,scheduled for elective fiberoptic bronchoscopy,were randomly assigned into 3 groups (n =28 each):normal saline group (group C),remifentanil infused at 3 ng· kg-1 ·min-1 group (group R1) and remifentanil infused at 5 ng· kg-1 · min-1 group (group R2).Responses to LMA insertion were defined as body movement and/or bucking during insertion.The initial target plasma concentrations of propofol were 5.2,4.8 and 4.4 μg/ml in groups C,R1 and R2,respectively.The target plasma concentration of propofol was determined by up-and-down sequential allocation.Each time the target plasma concentration increased/decreased by 0.2μg/ml.EC50 and 95 ％ confidence interval of propofol blunting responses to LMA insertion were determined by probit method.Results EC50 (95 ％ confidence interval) of propofol was 5.03 (4.92-5.12) μg/ml,4.71 (4.58-4.84) μg/rnl and 4.46 (4.20-4.94) μg/ml in groups C,R1 and R2,respectively.There was no significant difference in EC50 of propofol between groups R1 and C (P ＞ 0.05).EC50 of propofol was significantly lower in group R2 than in groups C and R1 (P ＜ 0.05).Conclusion The infusion rate of remifentanil should not be lower than 5 ng· kg-1· min-1 when combined with propofol in pediatric patients undergoing fiberoptic bronchoscopy.

This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.

Objective This paper is intended for establishing quality standard of Ershiwuwei Shanhu pill. Methods The TLC was used to indentify Pyrethrum tatsienense, Sesami nigrum semen, Aquilariae lignum resinatum, Caryophmlli flos, Acori calami rhizoma, Chebulae fructus, Aucklandiae radix and Glycyrrhizae radix et rhizome. For hydroxysafflor yellow A, the HPLC system consisted of WondaSil-C18 (250 mm×4.6 mm, 5μm), methanol-acetonitrile- 0.7% phosphoric acid (26:2:72) as mobile phase, flow rate of 1.0 ml/min, detection wavelength at 403 nm. For liquiritin, the HPLC system consisted of WondaSil-C18 (250 mm×4.6 mm, 5μm), acetonitrile (A)-0.05% phosphoric acid (B) as mobile phase, flow rate of 1.0 ml/min, detection wavelength at 237 nm. And for crocin-I, the HPLC system consisted of WondaSil-C18 (250 mm×4.6 mm, 5μm), methanol-water (49:51) as mobile phase, flow rate of 1.0 ml/min, detection wavelength at 440 nm. Results The methods of TLC were simple with strong specificity and good reproducibility. The results of HPLC showed that calibration curve was linear in the ranges of 0.046 7-0.233 8μg for hydroxysafflor yellow A, and 0.510 6-1.531 8μg for liquiritin and 0.048 1-0.340 5μg for crocin-I. The average recovery rate was 102.01%, 99.50%and 99.32%, respectively. Conclusion The new method is more appropriate for the quality control of Ershiwuwei Shanhu pill.

This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.
Actins ; genetics ; metabolism ; Animals ; Artemisia ; chemistry ; Artemisinins ; isolation & purification ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Epithelial Cells ; cytology ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; Idiopathic Pulmonary Fibrosis ; pathology ; Plants, Medicinal ; chemistry ; Pulmonary Alveoli ; cytology ; RNA, Messenger ; metabolism ; Rats ; Smad3 Protein ; genetics ; metabolism ; Smad7 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; pharmacology ; Vimentin ; genetics ; metabolism

OBJECTIVETo identify HLA-A0201 restricted cytotoxic T lymphocyte (CTL) epitopes derived from the hepatitis B virus e (HBe) antigen, for future use in a specific immunotherapy based on the identified epitope(s).

METHODSHBe gene sequences from the hepatitis B virus serotypes with the highest frequencies in China were analyzed by bioinformatic web-based interfaces for quantitative motif prediction, extended motif prediction, and peptide super-motif prediction. Four candidate peptides were identified: HBe1, HBe2, HBe3, and HBe4. The affinities of each were tested in vitro with T2 cells, which lack the transporter-associated with antigen transport (TAP) protein but express low levels of the MHC class I surface molecule, and measured by the T2 binding assay and DC50 assay. Flow cytometry was used to detect the fluorescence index of control and experimental groups.

RESULTSThe peptides HBe1 (LLWFHISCL), HBe2 (YLVSFGVWI), HBe3 (CLTFGRETV), and HBe4 (DLLDTASAL) were identified and tested as candidate targets. HBe2 and HBe3 showed higher HLA-A0201 affinity. HBe1, HBe2, and HBe3 showed better binding stability.

CONCLUSIONTwo peptides based on HBe antigen, YLVSFGVWI and CLTFGRETV, possess both sufficient binding affinity and stability and may represent useful HLA-A0201-restricted CTL epitopes. Further study is needed to determine the immunogenic properties of these two peptides in vivo.

Amino Acid Sequence ; Epitopes, T-Lymphocyte ; Hepatitis B e Antigens ; Hepatitis B virus ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.

METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.

RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).

CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.

Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics

China ; epidemiology ; Colorectal Neoplasms ; epidemiology ; genetics ; Folic Acid ; blood ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Incidence ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Polymorphism, Genetic ; Risk Factors

Objective To investigate the effect of recombinant murine interleukin-23(rIL-23)on systemic candidiasis in a murine model.Methods A cyclophosphamide-induced immunosuppressed murine model of systemic candidiasis was established.The mice were divided into control group and rIL-23 treatment group.Colony forming units(CFU)of the kidney and spleen were determined by using plating dilution method.The histopathological changes and degree of infection of the kidney and spleen were graded.Meanwhile,the levels of interferon gamma(IFN-γ)in the spleen were measured by enzyme-linked immunosorbent assay.Results On the 2nd,3rd and 7th day after Candida albicans infection the number of CFU of the fungi in the kidney in the control group was significantly greater than that in rIL-23 treatment group(P＜0.01).The number of CFU of the fungi on the 2nd,3rd and 7th day after Candida albicans infection in the spleen in control group was also greater than that in rIL-23 treatment group,but without statistically significant difference(P＞0.05).The scores of histopathological changes in the kidney in rIL-23 treatment group were lower than those in control group(P＜0.01),and the degree of infection was milder in rIL-23 treatment group.The scores of histopathological changes in the spleen in rIL-23 treatment group were also lower than those in control group,but without statistically significant difference(P＞0.05).The levels of IFN-γ in the spleen on the 2nd,3rd and 7th day after infection in rIL-23 treatment group were significantly higher than those in control group(P＜0.01).Conclusion rIL-23 has protective effect on murine systemic candidiasis.