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ABSTRACT: Wilms' tumor (nephroblastoma) is a cancer of the kidneys that occurs typically in children and rarely in adults. Early diagnosis is very important for the treatment and prognosis of the disease. The aim of our study was to discover and identify potential non-invasive and convenient biomarkers for the diagnosis of Wilms' tumor.
Nude mice were used to construct a Wilms' tumor model by injecting nephroblastoma cells into their bilateral abdomen. We collected 94 serum samples from mice consisting of 45 samples with Wilms' tumor and 49 controls. The serum proteomic profiles of the samples were analyzed via surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. The candidate biomarkers were purified by high-performance liquid chromatography, identified by liquid chromatography-mass spectrometry, and validated using ProteinChip immunoassays.
We finally retrieved two differential proteins (m/z 4509.2; 6207.9), which were identified as apolipoprotein A-II and polyubiquitin, respectively. The expression of apolipoprotein A-II was higher in the Wilms' tumor group than in the control group (P < 0.01). By contrast, the expression of polyubiquitin was lower in the Wilms' tumor group than in the control group.
Apolipoprotein A-II and polyubiquitin may be used as potential biomarkers for nephroblastoma in children, and the analysis of apolipoprotein A-II may help diagnose and treat Wilms' tumor.
Chinese medical journal 05/2012; 125(10):1727-32. · 0.86 Impact Factor
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ABSTRACT: Neuroblastoma (NB) is one of the most common malignant solid tumors of childhood. It is still not clear whether the apoptosis of tumor cells or the non-tumor cells contributes to the increase of concentration of cytochrome c (Cyt c) in the serum of the cancer patients. The aim of this research was to identify the source of the Cyt c in the serum when the tumor grows up by subcutaneous inoculation of human NB cells into nude mice.
We subcutaneously inoculated human NB cells (KP-N-NS) into nude mice and collected the sera of tumor-bearing mice (n = 14) and control mice (n = 25) 4 weeks later in order to screen for and identify differentially expressed proteins in the serum. Differentially expressed proteins in the serum were screened by surface-enhanced laser desorption/ionization-time-of-flight (SELDI-TOF) mass spectrometry.
The relative intensity of a protein having a mass-to-charge ratio (m/z) of 11 609 was 3338.37 ± 3410.85 in the tumor group and 59.84 ± 40.74 in the control group, indicating that the expression level of this protein in the tumor group was 55.8 times higher than that in the control group. Serum proteins were separated and purified by high-performance liquid chromatography (HPLC). Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was performed to produce peptide mass fingerprints (PMFs). Spectrum analysis and a database search revealed that the highly expressed protein (m/z = 11 605.4) from the serum of tumor-bearing mice was the mouse Cyt c.
Increased concentration of Cyt c in the serum of tumor-bearing nude mice might be partially attributed to the secretion of this protein by non-tumor cells.
Chinese medical journal 01/2012; 125(2):316-20. · 0.86 Impact Factor
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ABSTRACT: Angiogenesis and lymphogenesis which were promoted by vascular endothelial growth factor (VEGF) and VEGF-C are important in the growth and metastasis of solid tumors. The high level of VEGF and VEGF-C were distributed in numerous types of cancers, but their distribution and expression in Wilms tumor, the most common pediatric tumor of the kidney, was unclear.
To learn about the distribution, mass spectroscopy and immunohistochemistry were used to measure the level of VEGF and VEGF-C in serum and tissue of Wilms tumor.
The expression level of VEGF in serum of Wilms tumor was the same as in pre-surgery and control, so it was the same case of VEGF-C. Both of these factors were chiefly located in Wilms tumor tissue, but not in borderline and normal. In addition, the higher clinical staging and histopathologic grading were important elements in high expression of VEGF and VEGF-C. Gender, age and the size of tumor have not certainly been implicated in expression level of VEGF and VEGF-C.
The lymph node metastasis and growth of tumors resulted from angiogenesis and lymphogenesis which were promoted by VEGF and VEGF-C in Wilms tumor. The autocrine and paracrine process of VEGF and VEGF-C were the principal contributor to specific tissues of Wilms tumor but not to the entire body.
Chinese medical journal 11/2011; 124(22):3716-20. · 0.86 Impact Factor
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ABSTRACT: single-stage transanal endorectal pull-through (TEPT) is a new technique for surgery of Hirschsprung's disease (HD). TEPT can be assisted by laparoscopy (laparoscopic assisted transanal pull-through, LATP) or with non-additional procedure (total transanal endorectal pull-through, TTEP). This study was undertaken to evaluate the long-term outcome of these approaches in children with HD.
we retrospectively studied 131 patients (112 males and 19 females) aged 7 days to 14 years who underwent single-stage TEPT from October 2003 to July 2008. The medical records were reviewed for pre-, intra- and immediate post-operative complications. The data on stool pattern and complications were collected during the follow-up. Outcome was measured by continence evaluation score.
no patients had intraoperative complications, but 5 had minor immediate postoperative complications. Late postoperative complications in 12 patients included enterocolitis (4 patients, one with severe enterocolitis died 7 months after operation), soiling (6) and constipation (2). There was a significantly higher frequency of stool in patients aged more than 36 months and those with a resected colon more than 30 cm (P<0.05). LATP showed significantly higher frequency of stool and soiling (P<0.05). Of the 54 patients who were older than 3 years at the time of follow-up, continence score was normal in 10, good in 39, fair in 3, and poor in 2. Seventy-seven patients achieved good bowel control in 12.8 ± 8.11 months after operation, 93.5 5% of whom within 24 months. Stool function was not improved in patients more than 30 months old after operation.
the long-term outcome of single stage TEPT was excellent. There were few postoperative complications, and stool pattern improved gradually to an excellent level within 24 months. Internal plication can be a good option for reducing the dilated proximal colon.
World Journal of Pediatrics 02/2011; 7(1):65-9. · 1.22 Impact Factor
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ABSTRACT: To study the effect of simultaneously increasing PTEN gene expression and inhibiting Livin gene expression on the gastric carcinoma cell line (BGC823) and construct a recombinant vector expressing PTEN while simultaneously silencing Livin.
The siRNA expression unit against Livin gene (siLivin) was cleaved from pRNAT-U6.1-Livin vector and then inserted into pCL-neo-PTE to construct the recombinant vector pCL-neo-PTEN-siLivin. Then pCL-neo-PTEN-siLivinp, pCL-neo-PTEN, pRNAT-U6.1-Livin, pCL-neo and pRNAT-U6.1 were respectively transfected into the gastric carcinoma cell line (BGC823) with LipofectAMINE(TM) 2000. The mRNA and protein expression level of PTEN and Livin genes in each cell group was detected by fluorescent quantitative RT-PCR and Western blot.
Recombinant vectors of pCL-neo-PTEN, pRNAT-U6.1-Livin and pCL-neo-PTEN-siLivin were constructed successfully. After transfection with pCL-neo-PTEN-siLivin, the mRNA and protein expression level of PTEN (0.897±0.112) rose in BGC823 cells while Livin gene became silenced. And the characterization of regulated cell bioactivity improved. There were significant differences between transfected and control groups (P<0.05). And the inhibiting effect on the proliferation and metastasis of BGC823 cell by increasing PTEN expression and silencing Livin simultaneously was better than that only by regulating PTEN genes or Livin genes alternatively.
The recombinant vector of expressing PTEN and silencing Livin gene simultaneously is successfully constructed.
Zhonghua yi xue za zhi 09/2010; 90(34):2428-32.
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ABSTRACT: To explore the microRNA (miRNA) differential expression profile between nephroblastoma cell line G401 and normal embryonic kidney cell line CCC-HEK-1 so as to provide rationales for the role of miRNA in the pathogenesis of nephroblastoma.
Three samples from G401 cell line and another 3 samples from CCC-HEK-1 cell line were chosen as the experimental group and the control group respectively. miRNA profiles in these samples were analyzed by microarray. The threshold value used to screen up and down-regulated miRNAs was fold change ≥ 2 or ≤ 50.0%. And real-time PCR was used to validate the reliability of microarray.
Seventy-one miRNAs were found up-regulated in the experimental group and 11 of them were more than 8 times versus control group. In addition, 59 miRNAs were found down-regulated and 11 of them were 12.5% versus control group. Overall, there were significant differences in the expression of miRNA between G401 and CCC-HEK-1 cell lines.
There is a differential expression of miRNA between G401 and CCC-HEK-1 cell lines. It may be related with occurrence and metastasis of nephroblastoma.
Zhonghua yi xue za zhi 07/2010; 90(26):1845-8.
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Chinese medical journal 07/2010; 123(14):1939-41. · 0.86 Impact Factor
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ABSTRACT: Dendritic cells (DCs) are one of the most important antigen presenting cells in the human body, and DCs at various stages of maturation possess different or even opposite functions. The aim of this study was to investigate the influence of growth hormones on the functional status of cord blood-derived DCs encompassing immunophenotype, ability to excrete interleukin (IL)-12 and provoke autologous leukomonocyte.
Mononuclear cells were isolated from fresh cord blood, with IL-4 and granulocyte-macrophage colony-stimulating factor (GM-CSF) used to induce and stimulate the mononuclear cells. Growth hormone at different concentrations was used to modify DCs, and then DCs morphology, number and growth status were observed. The immunophenotype of DCs was detected with a flow cytometer. The concentration of IL-12 in the DCs supernatant was determined by enzyme linked immunosorbent assay (ELISA) and DCs functional status was evaluated by autologous mixed lymphocyte reactions.
Mononuclear cells from cord blood can be differentiated into DCs by cytokine induction and growth hormone modification. With the increase in growth hormone concentrations (5 - 100 microg/L), the expression of DCs HLA-DR, CD1alpha, CD80 and CD83 were significantly increased (P < 0.05). The ability of DCs to secrete IL-12 was significantly improved (P < 0.05), and the ability of DCs to activate autologous lymphocytes was significantly enhanced (P < 0.05). Pegvisomant was able to ablate the effects of growth hormone on DCs.
Growth hormone may facilitate DCs induction and maturation, and improve the reproductive activity of autologous lymphocytes in a dose-dependent manner. Growth hormone may serve as a factor of modifying DCs to achieving maturity.
Chinese medical journal 04/2010; 123(8):1078-83. · 0.86 Impact Factor
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ABSTRACT: Non-heart-beating donor lung has been a promising source of lung transplantation. Many studies on non-heart-beating donor lungs are based on animal lung transplantation. In this study, we assessed by organ bath the effect of one-hour warm ischemia on the non-heart-beating donor lung in terms of the integrity of contractile and relaxant functions and tissue structures of pulmonic arteries and bronchi.
Sixteen Swedish pigs were randomly classified into two groups: heart-beating donor group and 1-hour warm ischemia non-heart-beating donor group. Pulmonic and bronchial rings were taken from the isolated left lungs of the pigs. The pulmonic rings were stimulated by U-46619 (5.7 mol/L) and acetylcholine (10(-4) mmol/L) to assess the contractile abilities of smooth muscle and the endothelium-dependent relaxation response, respectively. As such, acetylcholine (10(-5) mmol/L) and natrium arachidonic acid (0.01%) were used to detect the contraction of bronchial smooth muscle and epithelium-dependent relaxation response. Meanwhile, the variances of precontraction tension of control groups were recorded to measure whether there was spontaneous relaxation during endothelium/epithelium-dependent relaxation course. Finally, papaverine solution (10(-4) mmol/L) was used to detect the non-endothelium/epithelium-dependent relaxant abilities of pulmonic and bronchial smooth muscles.
There was no significant difference in the tension values of precontraction of pulmonic rings (P > 0.05), endothelium-dependent relaxation (P > 0.05), precontraction of bronchial rings (P > 0.05) and epithelium-dependent relaxation (P > 0.05) between the heart-beating donor group and the 1-hour warm ischemia non-heart-beating donor group. And the pulmonic and bronchial rings of each subgroup B had no spontaneous relaxation. Finally, papaverine solution relaxed the smooth muscle of all the rings completely.
The results of this experiment suggest that the contractile and relaxant functions and tissue structures of pulmonic arteries and bronchi are not damaged after warm ischemia for 1 hour, and support the further study of non-heart-beating donor lung.
Chinese medical journal 12/2009; 122(23):2903-6. · 0.86 Impact Factor
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ABSTRACT: Dendritic cells (DCs) are the most important antigen-presenting cells in the human body, and DCs with different mature status possess different or even opposite functions. This study was designed to explore the influence of insulin on the functional status of cord blood-derived DCs and on DC-induced cytotoxic T lymphocyte (CTL) activity against pancreatic cancer cell lines.
Mononuclear cells were isolated from fresh cord blood. Interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) were used to induce or stimulate the mononuclear cells. Insulin at different concentrations served to modify DCs, and then DC morphology, number, and growth status were assessed. The DC immunophenotype was detected with a flow cytometer. The IL-12 in DC supernatant was determined by ELISA. DC functional status was evaluated by the autologous mixed lymphocyte reaction. T lymphocytes were induced by insulin-modified DCs to become CTLs. The CTL cytotoxicity against pancreatic cancer cell lines was determined.
Mononuclear cells from cord blood can be differentiated into DCs by cytokine induction and insulin modification. With the increase in insulin concentration (2.5-25 mg/L), the expression of DC HLA-DR, CD1alpha, CD80, and CD83 was significantly increased, the DC ability to secrete IL-12 was significantly improved, DC function to activate autologous lymphocytes was significantly enhanced, and the cytotoxicity of CTLs induced by insulin-modified DCs against pancreatic cancer cell lines was significantly strengthened.
Insulin may facilitate DC induction and maturation, and improve the reproductive activity of autologous lymphocytes. The cytotoxicity of CTLs induced by insulin-modified DCs against pancreatic cancer cell lines was significantly enhanced. Insulin may serve as a factor modifying DCs and inducing CTLs in vitro in insulin biotherapy.
Hepatobiliary & pancreatic diseases international: HBPD INT 10/2009; 8(5):529-34. · 1.08 Impact Factor
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ABSTRACT: Alterations in DNA methylation occur during the pathogenesis of human tumors. In this study, we investigated the influence of DNA methyltransferase 3b (DNMT3b) on fragile histidine trial (FHIT) expression and on DNA methylation of the FHIT promoter region in the hepatoma cell line SMMC-7721.
DNMT3b siRNA was used to down-regulate DNMT3b expression. DNMT3b and FHIT proteins were determined by Western blotting. Methylation-specific PCR was used to analyze the methylation status of the FHIT gene.
After DNMT3b siRNA transfection, the expression of DNMT3b was inhibited in SMMC-7721 cells, and the expression of FHIT was significantly higher than that in the control group. There was no significant difference in methylation status between the DNMT3b siRNA transfected cells and control cells.
DNMT3b may play an important role in regulation of FHIT expression in hepatoma SMMC-7721 cells, but not through methylation of the FHIT promoter.
Hepatobiliary & pancreatic diseases international: HBPD INT 07/2009; 8(3):273-7. · 1.08 Impact Factor
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ABSTRACT: To screen and characterize the serum protein biomarkers in nephroblastoma so as to establish the proteins as the specific serum biomarkers for diagnosis and prognosis monitoring.
The differential protein peaks were located by detecting serum samples of preoperative and postoperative patients and normal children using the SELDI-TOF MS technology. After purification, the differential proteins were further analyzed by LC-MS/MS and the protein sequences searched in database.
Two peaks with m/z of 6455.5 and 6984.4 were selected as potential biomarkers. They were weakly expressed in nephroblastoma (intensity: 1029 +/- 364, 297 +/- 126) but highly expressed in normal individuals (2108 +/- 837, 753 +/- 226); another peak with m/z of 9190.8 was weakly expressed in preoperative sera (283 +/- 154) but highly expressed in serum samples of postoperative patients and normal children (5974 +/- 657, 6231 +/- 519). The protein at 6455.5 and 9190.8 were identified as apolipoprotein C-III and haptoglobin respectively.
The detection of differentially expressed apolipoprotein C-III and haptoglobin may have potential utilities for serum diagnosis, malignancy classification and prognosis monitoring of nephroblastoma and is worthy of further studies and applications.
Zhonghua yi xue za zhi 06/2009; 89(18):1259-63.
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ABSTRACT: To detect and identify the potential specific serum biomarkers for diagnosis of papillary thyroid cancer.
Samples of 35 patients with papillary thyroid carcinoma, 40 patients with benign thyroid nodule and 34 healthy individuals were analyzed using the SELDI-TOF ProteinChip System and bioinfomation technology to find the differential peaks which were separated by HPLC and then further analyzed by LC-MS/MS. The protein sequences were analyzed by SEQUEST software and searched in Bioworks database.
The top six mass-to-charge ratio (M/Z) peaks with the smallest P value were 6651, 6452, 7653, 7932, 15 106 and 15 848 Da, respectively. The 6651 and 6452 Da proteins were weakly expressed in papillary thyroid carcinoma but highly expressed in benign thyroid nodules and healthy individuals. The differences had statistical significance (P < 0.01). The 7653, 7932, 15 106, 15 848 Da proteins were highly expressed in papillary thyroid carcinoma but weakly expressed in benign thyroid nodules and healthy individuals. The differences were statistically significant (P < 0.01). Combination of these six proteins, using the method of leave-one-out to make crossing detection, the specificity of discriminating papillary thyroid carcinoma and non-cancer was 88.0%, and its sensitivity was 92.5%. The 6651 and 6452 Da proteins were identified as apolipoprotein C-I and apolipoprotein C-III, respectively. The 7653 and 15 106 Da proteins were identified as the same protein-alpha-globin, and the 7932 and 15,848 Da proteins were identified as the same protein-beta-globin.
The detection of differentially expressed apolipoprotein C-I, apolipoprotein C-III, alpha-globin, and beta-globin may have utility for diagnosis of papillary thyroid carcinoma and are worthy of further investigation.
Zhonghua zhong liu za zhi [Chinese journal of oncology] 05/2009; 31(4):265-8.
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ABSTRACT: To detect the effect of functional status and immunophenotype of dendritic cell modified with growth hormone (GH) from by cord blood.
Monocyte was isolated from cord blood, DCs were induced with IL-4 & GM-CSF and modified with different concentration of GH. The morphology, count and growth status of DC were observed appearance. And the immunophenotype of DC was detected with flow cytometry. IL-12 in supernatant was measured with ELISA. The immuno-activity was determined with autologous mixed lymphocyte reaction.
Mature DC were induced from adherent lymphomonocyte under the action of IL-4, GM-CSF and GH of different densities (5, 25, 100 ng/ml). GH increased the positive expression of CD83 (31% +/- 4%, 34% +/- 4%, 34% +/- 5%), CD80 (26% +/- 4%, 29% +/- 6%, 30% +/- 6%)and HLA-DR (45% +/- 8%, 49% +/- 6%, 50% +/- 5%) (P < 0.05), enhanced the ability of secreting IL-12 (P < 0.05), and boosted significantly the immuno-activation of DC to autologous lymphocyte (P < 0.05).
GH could promote the differentiation, maturation and the expression of costimulatory molecules, increase the IL-12 production in DC and enhanced the ability of DC to stimulate the proliferation of lymphocyte within a certain dose range.
Zhonghua yi xue za zhi 04/2009; 89(16):1139-43.
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ABSTRACT: To investigate the influence of DNA methyltransferase (DNMT) 3b on the expression of cyclin D1 gene and methylation of its promoters and to investigate the function of DNMT3b.
Human hepatocellular carcinoma cells of the line SMMC7721 were cultured and randomly divided into 3 groups: experimental group transfected with siRNA to silence the DNMT3b, control group transfected with control siRNA, and normal group without transfection. The transfection rate of siRNA was detected by fluorescence microscopy. MTT method was used to measure the survival rate of the SMMC-7721 cells. Western blotting and cell proliferation assay were performed to evaluate the expression of cyclin D1 and cell growth. Methylation specific PCR (MSP) was performed to investigate whether the promoter of cyclin D1 was methylated.
Fluorescence microscopy showed that the transfection rate of siRNA was over 90%. MTT method showed that 24 h and 36 h after transfection the A value and survival rate of the SMMC7721 cells of the experimental group were both significantly higher than those of the control d normal groups (all P < 0.05). Western blotting showed that the expression levels of DNMT3b and cyclin D1 of the experimental group decreased significantly compared with the control and the normal groups. MSP showed no obvious change of the state of methylation among the 3 groups.
DNMT3b may regulate the expression and the function of cyclin D1 gene in the human hepatocellular carcinoma cells, but does not change its methylation state. DNMT3b may play their role as a signal transduction element rather than as a DNA methyltransferase.
Zhonghua yi xue za zhi 04/2009; 89(8):555-8.
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ABSTRACT: The objective of this study was to identify a specific fingerprint chromatogram model of serum proteins for early screening and diagnosis of Hirschsprung disease.
To detect the protein mass spectrograms of 78 serum specimens (42 specimens of Hirschsprung disease, 16 specimens of adhesive ileus including appendicitis and Meckel diverticulum after operation and inflammatory bowel disease, and 20 specimens of normal control subjects), we used surface-enhanced laser desorption/ionization time of flight mass spectrometry technology, combined with bioinformatics methods (support vector machine) to develop and compare protein mass spectrograms from serum samples.
We identified 3 protein markers, the mass-to-charge ratio of which is positioned at 3221.7, 5639.2, and 6884.2 from the fingerprint chromatogram model of serum protein for early screening and diagnosis of Hirschsprung disease. The markers had 100% sensitivity and specificity.
The fingerprint chromatogram model of serum protein using surface-enhanced laser desorption/ionization time of flight mass spectrometry technology combining support vector machine is a new method of early screening and diagnosis of Hirschsprung disease that is worthy of additional research and application.
PEDIATRICS 08/2007; 120(1):e56-60. · 4.47 Impact Factor
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ABSTRACT: To find new biomarkers and to establish serum protein fingerprint models for early detection and diagnosis of nephroblastoma by SELDI-TOF-MS and bioinformatics tools.
Seventy five serum samples from 35 nephroblastoma patients, 30 children's abdominal solid tumor patients, and 20 healthy children were bound to WCX2 protein chip and tested by surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS). The data of spectra were analyzed by support vector machine(SVM).
Four peaks with m/z of 6984.5, 6455.5, 6914.0, 3256.7 were selected as potential biomarkers. The detective model combined with 2 biomarkers could separate nephroblastoma from the healthy group with a sensitivity of 100%, and a specificity of 100%. The diagnostic model combined with 2 biomarkers could separate nephroblastoma from other child's abdominal solid tumors with a sensitivity of 93.3%, and a specificity of 100%.
High sensitivity and specificity achieved by this method show great potential for early diagnosis of nephroblastoma, and screening for new tumor biomarkers.
Zhonghua yi xue za zhi 12/2006; 86(42):2982-5.
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ABSTRACT: To find new biomarkers and establish serum protein fingerprint models for early diagnosis and preoperative staging of papillary thyroid carcinoma, we employed SELDI-TOF-MS and bioinformatics tools. A total of 116 samples were analyzed in this study. The first 80 samples were analyzed by SELDI-TOF-MS and two biomarker patterns were identified. Pattern 1 distinguishes patients with papillary thyroid carcinoma from healthy individuals. Pattern 2 distinguishes papillary thyroid carcinoma from benign thyroid nodes. The remaining 29 samples were analyzed on the second day and served as an independent test set. The analysis of this independent test set yielded a specificity of 80.0% and a sensitivity of 88.9% for pattern 1 and a specificity of 80.0% and a sensitivity of 80.0% for pattern 2. Two additional biomarker patterns were identified to distinguish different stages of the papillary thyroid carcinoma (pattern 3) with an accuracy of 77.1% and different pathological types of thyroid carcinoma (pattern 4) with an accuracy of 88.1%. Taken together, the SELDI-TOF-MS technique combined with bioinformatics approaches can not only facilitate the discovery of better biomarkers for papillary thyroid carcinoma but also provide a useful tool for molecular diagnosis in the future.
PROTEOMICS 11/2006; 6(19):5344-9. · 4.51 Impact Factor
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ABSTRACT: Dendritic cells (DCs) loaded with complex antigen are always used to induce cytotoxic T lymphocytes (CTLs) which have a specific anti-tumor activity. However, CTLs can assault autologous cells induced by DCs loaded with autologous antigen. This study aimed to explore how to weaken the autoimmune reaction induced by DC vaccine by combining mature DC (mDC) activating immunity and immature DC (imDC) leading to immune tolerance to make hepatocellular carcinoma (HCC) vaccine in vitro.
DC progenitors derived from human peripheral blood were assigned to two groups. One was cultured to mDC and pulsed with frozen-thawed antigen (FTA) of human HCC cell line SMMC-7721 cells (mDC group), and the other was cultured to imDC and pulsed with FTA of human liver cell line L-02 cells (imDC group). The morphology of DCs was monitored and cells phenotypes including HLA-DR, CD80, CD1alpha, CD83 were assayed by flowcytometry (FCM). The concentrations of interleukin-12 (IL-12) in the supernatant were assayed by ELISA. Methyl thiazolyl tetrazolium (MTT) was used to evaluate T cell proliferation induced by mDC and imDC and the killing rate of CTL induced by mDC and imDC respectively/together on SMMC-7721 and L-02 cells.
Compared with the imDC group, the mDC group was characterized by the following: increased secretion of IL-12 (P<0.05); higher expression of HLA-DR, CD1alpha, CD80, CD83; and stronger activity in stimulating proliferation of isogenic T cells (P<0.05). CTL induced by the mDC group had a significant killing response to SMMC-7721 as well as a higher killing rate for L-02 (P>0.05). CTL induced by mDC and imDC together had a higher killing response to SMMC-7721, but a lower killing rate for L-02 (P<0.01).
CTL induced by mDC and imDC together has a higher antigen-specific killing response in vitro than that induced by mDC alone. This may be of greater clinical value.
Hepatobiliary & pancreatic diseases international: HBPD INT 08/2006; 5(3):422-7. · 1.08 Impact Factor
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ABSTRACT: To detect new biomarkers and to establish a serum protein fingerprint model for early detection and diagnosis of thyroid cancer.
The serum samples of 40 thyroid cancer patients, 9 thyroid adenoma patient, and 32 healthy individuals were randomly divided into 2 sets: training set (n = 66, including 32 thyroid cancer patients, 9 thyroid adenoma patients, and 25 healthy individuals) and test set (n = 15). The serum protein were bound to WCX2 chip and tested by surface enhanced laser desorption/ionization time of flight-mass spectrometry (SELDI-TOF-MS). The data of spectra were analyzed by support vector machine (SVM) to establish a diagnostic model.
The detective model combined with 3 biomarkers could differentiate the serum of thyroid cancer from that of healthy individual with a specificity of 86% and a sensitivity of 100%. The diagnostic model combined with 3 biomarkers could differentiate thyroid cancer from thyroid adenoma with a specificity of 88.9% and a sensitivity of 96.9%. The positive predictive value to differentiate papillary thyroid carcinoma from the thyroid cancer of other types was 97%, and the positive predictive value of thyroid carcinoma of other pathological types was 71%.
The combination of SELDI with bioinformatics tools is a novel, effective, and highly specific and sensitive method for thyroid cancer detection and diagnosis.
Zhonghua yi xue za zhi 05/2006; 86(14):979-82.
