Objective:To study inhibited tecomerase activity of HL-60 cells in human cancer cell by Icarrin(ICA) and its mechanism.Methods:MTT assay,NBT assay, TRAP-PCR,RT-PCR assay,Flow Cytometry.Results:ICA inhibited telomerase activity in HL-60 cells significantly.It was negative correlation between telomerase activity and the expression ratio of CD11b antigen in HL-60 cells.It can induced HL-60 cells to differentiate into mature granulocytes.It can changed cell cycle distribution of HL-60 cells,which was reflected by the accumulation of the vast majority of cells in the G0/G1phase and the loss of cells in the S phase. It can downregulated cell proliferation related gene c-myc;upregulated p21 gene protein and mRNA expression.Conclusion:This study proved mechanism of ICA on inhibited telomerase activity from gene-protein-effect of cell level in HL-60 cells.

ObjectiveTo investigate the methylation status of multiple genes in plasma and tumor tissues and its application in molecular diagnosis of esophageal squamous cell carcinoma (ESCC).Methods Methylation specific polymerase chain reaction (MSP) was used to detect methylation status of 5 tumor suppressor genes,such as adenomatous polyposis coli ( APC ),retinoic acid receptor-beta2 ( RARβ2 ),E-cadherin (CDH1),cyclin-dependent kinase inhibitor4A (p16INK4α) and ras association domain family member 1 A (RASSF1A) in tumour tissues,adjacent normal tissues and plasma which obtained 1 d preoperative and 7 d postoperative in 76 cases with ESCC.60 healthy volunteers were randomly selected as a control which were age-matched and sex-matched.Chi square test was used to analyze DNA methylation rates of 5 genes in various groups of tissue and plasma samples; Kappa test was used to compare the consistency of DNA methylation in the plasma samples and tissue samples,and their correlation was analyzed by Spearman correlation test; Receiver operating characteristic curve (ROC) was used to evaluate the sensitivity and specificity for single gene detection and 5 genes joint detection for diagnosis of esophageal squamous cell carcinoma.ResultsThe methylation rates of APC,RARβ2,CDH1,p16INK4α and RASSF1A in tumour tissues of patients with ESCC were 44.7％ ( 34/76),72.4％ ( 55/76 ),72.4％ (55/76),86.8％ ( 66/76 ),55.3％ (42/76),respectively,which were significantly higher than that in the corresponding adjacent normal tissues [ 6.6％ ( 5/76 ),3.9％ ( 3/76 ),3.9％ ( 3/76 ),3.9％ ( 3/76 ),2.6％ ( 2/76 ),x2 =29.01,75.39,75.39,105.34,57.18,all P ＜ 0.001 ].The methylation rates of above 5 genes in patients' plasma were 42.1％ ( 32/76 ),63.2％ ( 48/76 ),63.2％ ( 48/76 ),71.1％ ( 54/76 ),50.0％ ( 38/76 ),respectively,which were significantly higher than that of control group [3.3％ (2/60),3.3％ (2/60),1.7％ ( 1/60),3.3％ (2/60),1.7％ (1/60),x2 =26.88,51.62,55.01,63.48,38.30,all P ＜ 0.001 ].The methylation consistency was favorable or well between plasma and tumour tissues in patients with ESCC ( Kappa value was 0.679,0.791,0.791,0.542 and 0.895,respectively.all P ＜0.001 ).In single-gene detection for patients' plasma,methylation,the sensitivity of 5 genes was 42.1％ ( 32/76 ),63.2％ ( 48/76 ),63.2％ ( 48/76 ),71.1％ ( 54/76 ),50.0％ ( 38/76 ),respectively.The specificity was 96.7％ ( 58/60 ),96.7％ ( 58/60 ),98.3％ (59/60),96.7％ (58/60),98.3％ (59/60),respectively.The area under curve (AUC) of ROC was 0.694 [95％ confidence interval( CI)0.606 - 0.782 ) ],0.799 ( 95％ CI 0.723 - 0.875 ),0.807 ( 95％CI 0.733 - 0.882),0.839 ( 95 ％ CI 0.769 - 0.908 ) and 0.742 ( 95 ％ CI 0.659 - 0.824 ),respectively.In united testing of 5 genes,the sensitivity was 80.3％ and the specificity was 88.3％,AUC was 0.843 (95％CI 0.773 -0.913 ).The sensitivity of united testing was significantly higher than that of single-gene detection of APC and RASSF1A(x2 =23.30,15.33 ; P ＜ 0.001 ),except RARβ2,CDH1 and p16INK4α (x2 =5.48,5.48,1.75; P =0.019,0.019,0.186);There was no significant differences in specificity between united testing and single-gene detection (x2 =1.922,1.922,3.348,1.922,3.348,all P ＞ 0.05 ).Conclusions The methylation consistency is favorable or well between tumour tissues and plasma in patients with ESCC.There is no significant superior in diagnosing ESCC with united testing of multiple tumor suppressor genes methylation in plasma than with single-gene detection.But the sensitivity of the former is better than the latter.

Objective To detect the expressions of microRNA-126 (miR-126) and microRNA-7 (miR-7) in esophageal squamous cell carcinoma (ESCC) and to analyze their correlations with clinicopathologic features and prognosis of ESCC.Methods The expressions of miR-126 and miR-7 in 116 ESCC samples and matched normal tissue samples were detected by real-time PCR.Statistical analysis was used to find the relationships among the expressions of miR-126 and miR-7,pathological characteristics and prognosis.Results Low expression,normal expression and high expression of miR-126 were found in 73 (62.9％),35 (30.2％) and 8 (6.9％) carcinoma samples respectively.Low expression,normal expression and high expression of miR-7 were found in 52 (44.8％),35 (30.2％) and 29 (25.0％) carcinoma samples respectively.The disease-free survival in patients with low expression of miR-126 and miR-7 was shorter than that in patients with non-low expression (x2 =4.268,P ＜0.05 ; x2 =4.993,P ＜0.05).The low expression of miR-126 was correlated with tumor location,family history and drinking (x2 =14.564,P ＜ 0.05 ; x2 =5.691,P ＜ 0.05 ; x2 =4.971,P ＜ 0.05),but was uncorrelated with gender,age,diferentiation,infiltration,lymphatic metastasis and smoking (all P ＞ 0.05).The low expression of miR-7 was uncorrelated with pathological characteristics of ESCC (all P ＞ 0.05).Conclusion The low expressions of miR-126 and miR-7 may be related to the prognosis of patients with ESCC,and have a certain clinical detection significance.

Objective To investigate the diagnostic and therapeutic experience in children with ulcerative colitis(UC) from clinical data.Met-hods The literature on UC in children publicated from 1995 to 2004 were retrieved by CBM,CNKI and VIP database.The clinical manifestations,X-ray,endoscopic and histologic features,management and prognosis of 172 patients in this hospital correspond with the inclusion criteria were retrospectively investigated and a definite diagnosis with UC was made.Results Of 172 patients,the ratio of maleold female was 1.07:1.0,and the age of patients ranged from 2 months to 15 years old.Thirty-six patients(36.7%)were less than 3 years-old;the course ranged from 3 days to 4 years.One patient had family history.The main manifestations were chronic diarrhea,mucus bloody stool,bloody purulent stool and abdominal pain.The systemic symptoms were fever,weight loss,malnutrition and anemia,no enteral symptoms and complications.The lesions were mainly seen in the whole colon with moderate to severe degree.The therapies based on salicylaxosulfapyridine(SASP) or 5-acetylsalicylic acid(5-ASA) together with corticosteroid in the short period have clinical remission in different levels.From long follow-up,the complete remittence was less than one third.Conclusions It is not rare to see UC in infantile.The clinical characteristics of UC in children are different from those in adults,the same is true before and after 3 year-old.The UC in children has high misdiagnosis and is difficult to manage.It is very necessary to establish a diagnostic and therapeutic guideline for children with UC in China.

Objective: To investigate the influence of Icarrin(ICA) on transforming growth factor ?2(TGF?2) expression by PG cells and the mechanisms of reversion of TGF?2 mediated immunosuppression by ICA.Methods: Cell proliferation and cytotoxici- ty were assayed by MTT assay. mRNA level of TGF?2 and perforin was detected by RT-PCR assay. assay.Expression of TGF?2 and IL- 2R? was analysed by folw cytometry assay. Results: ICA could inhibit the protein and mRNA expression of TGF?2 in PG cells. It could also reverse the supressed cytotoxicity of LAK, CD3AK cells by TGF?2. The IL-2R? expression on LAK and perforin gene transcription and proliferation of CD3AK inhibited by TGF?2 were also partly recovered. Conclusion: The anti-anti-tumor activity of ICA is at least partly due to its capability of inhibiting TGF?2 expression in tumor cells and restoring the cytotoxicity of immunocompetent cells inhibited by TGF?2.

Objective To investigate the existence of HBV DNA YMDD mutation in patients with chronic hepatitis B and the related significance. Methods HBV DNA YMDD mutation was detected in 90 patients with hepatitis B. The liver function, HbeAg, HBVDNA, and HBV DNA YMDD mutation in patients with HBV DNA YMDD mutation were analyzed. Results HBV DNA YMDD mutation was found in 20 out of 90 patients with hepatitis B (positive rate 12.00%). HBV DNA YMDD mutation was found in 9 out of 36 patients receiving lamivuding therapy (positive rate 25.00%), but HBV DNA YMDD mutation was found in 11 out of 54 patients without lamivuding therapy (positive rate 20.37%). In the 20 patients with HBV DNA YMDD mutation, 5 patients were negative in HBeAg. HBV DNA, ALT, and Tb of the 20 patients with HBV DNA YMDD mutation were 3.0221?10 5-2.369?10 8 (5.41?7.75?10 7) copies/ml, 34 -1 216 (288.20?338.07) IU/L, and 8.9-326.7 (48.74?72.20) ?mol/L, respectively. Conclusion HBV DNA YMDD mutation may occur during the process of lamivuding therapy or may exist naturally. YMDD mutation may co-exist with pre-C gene mutation, suggesting that mutation often occurs in many sites in HBV genes. The reproduction level of HBV DNA ranges from the middle to the high levels. The pathogenicity of YMDD mutation species is various. The liver function may be normal and may also be abnormal in various degrees in these patients with HBV DNA YMDD mutation.

Objective To explore the clinical effect of mycobacterium tuberculosis determination by culture and microscopy assay from sputum specimens of pulmonary tuberculosis patients.Methods The sputum specimens were dyed with the Ziehl-Neelsen anti-acid dyeing direct microscopy and examined by BACTEC-TB 960 culture system.Results The positive rate of culture and microscopy were 36.6% and 24.6%,the difference in the positive rate between the two methods was significant(P

AIM Mechanism of icariin (ICA) and pseudomonas jinanensis agent (PJA) on anti-metasta-sis of a highly metastatic human lung tumor cell line (PG) and the effect type of the combination of ICA,PJA were studied. METHODS PG cells were treated with ICA、PJA, or the combination of both in vitro. Adhesion assay, invasion assay, migration assay, flow cytometry (FCM) and immunohistochemical straining were used. The effects of ICA and PJA combination were evaluated with Q method of Jin Zheng-Jun. RESULTS ICA、PJA could decrease PG cells adhesive ratio to laminin substrate in a time-dependent manner ( P

Human CD3AK cells were prepared from peripheral blood mononuclear cells by culturing with recombinant IL-2 and antiCD3AK McAb. The mechanism and regulation of CD3AK cytotoxic activity with cytokines (rhIFN-?, rhIFN-?, TNF) and chernotherapeutic agents (CDDP or ADM) were observed by LDH-release assay, ABC-CELISA and the flow cytometric assay. The results showed: (1) Adhesion molecules ICAM-l/LFA-1 participated in CD3AK-mediated killing of tumor cells, hrlFN-? and TNF enhanced cytotoxicity of CD3AK through this pathway. (2) CD3AK could indirectly kill tumor cells by releasing soluable cytotoxic factors. (3) The membrane-associated TNF may be involved in CD3AK-mediated cytotoxicity. (4) CD3AK cells could induce the apoptosis of tumor cells. (5) Pretreatment of tumor cells with CDDP or ADM resulted in the increased vulnerability of tumor cells to CD3AK-mediated killing, the enhancement of CD3AK-mediated cytotoxicity by CDDP was relative to the increased expression of ICAM-1, HLA-ABC on tumor cell membrane.

OBJECTIVETo explore if the antileukemic drugs Vp16 or Ara-C are able to upregulate DR5 gene expression and enhance Apo2L-induced apoptosis of HL-60 cells.

METHODSCell apoptosis was determined by flow cytometry after annexin V/PI staining, the effect of Apo2L on fresh leukemia cells by MTT reduction assay, the expression of DR5 gene in HL-60 cells by semi-quantitative RT-PCR.

RESULTS1. Apo2L induced apoptosis of HL-60 cells in a dose-dependent manner. 2. Apo2L inhibited the proliferation of fresh leukemia cells, but there was difference among different subtypes. 3. Vp16 or Ara-C upregulated DR5 gene expression and augmented Apo2L-induced apoptosis in HL-60 cells.

CONCLUSIONApo2L could induce apoptosis of HL-60 cells and inhibit the proliferation of fresh leukemia cells. Ara-C or Vp16 upregulated DR5 gene expression and increased the sensitivity of HL-60 to Apo2L-induced cytotoxicity. Apo2L might be a promising antileukemic agent for the treatment of leukemia.

Antineoplastic Agents ; pharmacology ; Apoptosis Regulatory Proteins ; Cytarabine ; pharmacology ; Drug Synergism ; Etoposide ; pharmacology ; Female ; HL-60 Cells ; Humans ; Leukemia ; drug therapy ; Male ; Membrane Glycoproteins ; pharmacology ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; Receptors, Tumor Necrosis Factor ; genetics ; TNF-Related Apoptosis-Inducing Ligand ; Tumor Necrosis Factor-alpha ; pharmacology