Objective To explore the microRNA （miRNA） expression changes and related miRNA characteristics of colorectal cancer （CRC） with hepatic metastasis by miRNA microarray. Methods The fresh specimens of primary CRC were collected in 10 patients during operation， which with hepatic metastasis or not. miRNA microarray analysis was performed to compare the miRNA expression levels in two groups. The different expression levels of miRNA were validated by quantitative real-time PCR analysis. Results A total of six dysregulated miRNAs were identified in the CRC patients with hepatic metastasis comparing with CRC patients without hepatic metastasis， including 3 up-regulated miRNAs （miR-224， miR-1236， and miR-622） and 3 down-regulated miRNAs （miR-155， miR-342-5p， and miR-363）， and the quantitative real-time PCR result of miR-224 consisted with the microarray finding. Conclusions miR-224 may be involved in the process of CRC with hepatic metastasis pathogenesis. miR-224 would be a research direction on a new biomarker or therapic method in CRC with hepatic metastasis.
ObjectiveTo screen for the differentially expressed genes in steroid-induced osteonecrosis of the femoral head (ONFH) by gene microarray.
MethodsThe femoral head tissue of ONFH was harvested from 3 patients with steroid-induced ONFH, aged 25, 31, and 38 years, respectively. Normal tissue was harvested from a 26-year-old male remains contributor. HE staining of the specimens was performed for observing the histology manifestation; the total RNA was extracted for measuring the purity; cDNA probe was synthesized by reverse transcription, and then were hybridized as the cDNA microarray for scanning of fluorescent signals and differentially expressed genes in the tissues.
ResultsHE staining of normal tissue showed complete unit composed of lamellar bone, continuous and complete lamellar bone with a concentric arrangement around blood vessels, and normal bone cells in the trabecular bone lacuna. In ONFH tissue, adipose tissue increased in the medullary cavity, with increased fat cells filling in the medullary cavity and extruding capillary, and with decreased bone cells in the bone trabecula, which had deeply-stained nuclear chromatin, pyknotic or cracking nucleus, and even bone cells disappeared in the part of the bone lacuna, and trabecular bone became thin, sparse, interrupt, reduced area in visual field/unit. Total RNA extraction electrophoretogram displayed clear bands of 28S and 18S, and the brightness ratio of the 28S:18S was 2:1, indicating good total RNA quality. And 44 genes were differentially expressed, and there were 28 up-regulated genes and 16 down-regulated genes, including cell/organism defense genes, cell structure/motility genes, cell division genes, cell signaling/cell communication genes, cell metabolism genes, gene/protein expression genes, and unclassified genes.
ConclusionThe analysis of the gene expression profile of steroid-induced ONFH can provide evidence for the pathogenesis of ONFH.
Objective To investigate the role of KiSS-1 gene in the metastatic process of carcinoma of gallbladder and the clinicopathologic significance of KiSS-1 gene expression in carcinoma of gallbladder. Methods Pathological specimens from 59 gallbladder carcinoma tissues (13 hepatic invasion and 13 lymphatic invasion tissues were included), matched with 7 para-tumor and 6 normal gallbladder tissues, were examined for the expression of KiSS-1 gene by tissue microarray technique and immunohistochemistry (EnVision). Results The positive rate of KiSS-1 expression was down-regulated (P＜0.05) in tumor tissues, as compared with normal and para-tumor tissues. In carcinoma of gallbladder, the expression of KiSS-1 had no relationship with the gender, age, tumor size, histological grade or differentiation, and metastasis of lymph node, while was associated with the depth of infiltration, invasion of liver and the clinical stages (Nevin). In Ⅰ+Ⅱ, Ⅲ+Ⅳ and Ⅴ stage, the positive rates of KiSS-1 were 92.3%, 57.1% and 27.8% respectively, with an undeniably clear lowering tendency (P=0.002). Conclusion Down-regulating expression of KiSS-1 is closely associated with the processes of genesis, invasion and metastasis in carcinoma of gallbladder, and may participate in regulating these processes.
Objective To identify the expression of pleiotrophin （PTN） mRNA and protein in the colorectal cancer tissues， and to explore the clinical value of it. Methods The expressions of PTN mRNA and protein in colorectal cancer tissues （colorectal cancer group） as well as normal colorectal tissues （normal control group） were tested by using in-situ hybridization and immunohistochemistry. Results The positive rates of PTN mRNA 〔63.9% （53/83） vs. 40.7%（22/54）〕 and protein〔60.2%（50/83） vs. 33.3%（18/54）〕 in colorectal cancer group were all significantly higher than those of normal control group （P=0.008， P=0.002）. There were no significant relationship between expressions of PTN mRNA and protein with gender， age， and type of tumor （P＞0.05）， but in tissues of Ⅲ-Ⅳ stage and poor differentiation，the positive rates of PTN mRNA and protein were all higher （P＜0.05）. Conclusions The over expressions of PTN mRNA and protein in colorectal cancer tissues may directly related to the invasion and metastasis. Meanwhile， it can be used as an index to predict metastasis and prognosis of colorectal cancer.
Objective To seek for a method of constructing the tissue microarray which contains keloid, skin around keloid, and normal skin. Methods The specimens were gained from patients of voluntary donation between March and May2009, including the tissues of keloid (27 cases), skin around keloid (13 cases), and normal skin (27 cases). The specimens were imbedded by paraffin as donor blocks. The traditional method of constructing the tissue microarray and section were modified according to the histological characteristics of the keloid and skin tissue and the experimental requirement. The tissue cores were drilled from donor blocks and attached securely on the adhesive platform which was prepared. The adhesive platform with tissue cores in situ was placed into an imbedding mold, which then was preheated briefly. Paraffin at approximately 70℃ was injected to fill the mold and then cooled to room temperature. Then HE staining, immunohistochemistry staining were performed and the results were observed by microscope. Results The constructed tissue microarray block contained 67 cores as designed and displayed smooth surface with no crack. All the cores distributed regularly, had no disintegration or manifest shift. HE staining of tissue microarray section showed that all cores had equal thickness, distinct layer, manifest contradistinction, well-defined edge, and consistent with original pathological diagnosis. Immunohistochemistry staining results demonstrated that all cores contained enough tissue dose to apply group comparison. However, in tissue microarray which was made as traditional method, many cores missed and a few cores shifted obviously. Conclusion Applying modified method can successfully construct tissue microarray which is composed of keloid, skin around keloid, and normal skin. This tissue microarray will become an effective tool of researching the pathogenesis of keloid.
Aiming at long signal acquisition time, low flux, bad signal-to-noise ratio and low intelligence in coloration biochip reader, a new kind of rapid device with high flux was developed. The device consisted of hardware system and software system. It used a charge-coupled device (CCD) as the photoelectric sensor elements and obtained the biochip microarray image. The device integrated the embedded operating system based on i.MX6 chip. The microarray image processing, data analysis and result output were achieved through the code information of the software chip. Experiments with the standard grayscale sheet and standard format chip were carried out. The results showed that the maximum measurement error was less than 0.1%, the value of R2 was 98.7%, and the value of CV was 1.096 1%. The comparison results of 200 samples showed that detection performance of the proposed device was better than that of the same kind of marketed equipment.
Objective To study hepatocyte growth factor (HGF) and its receptor (c-met) expressions in human colorectal cancer and non-neoplasm colorectal mucosa, and the relationship with tumor angiogenesis. Methods Tissue microarrays (TMAs) were made up of 80 cases of colorectal cancer and 80 cases of nonneoplasm colorectal mucosa. The expressions of HGF and c-met were detected by immunohistochemistry (SP). CD105 was used as a marker to account microvessel density (MVD) in tumor tissue. Results HGF was over expressed in 48 cases and c-met was over expressed in 63 cases of colorectal cancer tissue, and the correlation between HGF and c-met positive expression was significant (r=0.231, Plt;0.05). The high expression rate of HGF and cmet in colorectal cancer were significantly higher than that in non-neoplasm colorectal mucosa (χ2=35.387, Plt;0.05; χ2=59.854, Plt;0.05) of colorectal cancer. The overexpression of HGF was correlated with lymph node metastasis (χ2=4.743, Plt;0.05) and TNM staging (χ2=5.576, Plt;0.05). The overexpression of c-met was correlated with differentiation (χ2=15.767, Plt;0.05) and lymph node metastasis (χ2=5.765, Plt;0.05) of colorectal cancer. MVD was different between overexpression and lowexpression colorectal cancer tissues of HGF and cmet (t=2.150, Plt;0.05; t=2.052, Plt;0.05). There was statistical correlation between HGF and cmet overexpression (r=0.259, Plt;0.05). The overexpressions of HGF and cmet were correlated with lymph metastasis in moderate differentiation cancer (χ2=13.154, Plt;0.05; χ2=5.371, Plt;0.05). Conclusions The overexpressions of HGF and c-met in colorectal cancer may be related with tumor angiogenesis. Detecting the expressions of HGF and c-met is valuable to estimate the biological character of colorectal cancer.
Objective To investigate the change of immunologic gene expression in cases of colorectal cancer with liver metastasis. Methods The total RNAs were extracted from tumor tissues of original lesions in 16 patients with colorectal cancer， DNA microarray was used to examine the change of immunologic gene expression in colorectal cancer patients with or without liver metastasis. Results Compared with samples without liver metastases， the expressions of 11 immunologic genes obviously down-regulated in the tumor tissues of colorectal cancer patients with liver metastasis， including：carboxypeptidase D；Fc fragment of IgE， high affinityⅠreceptor for gamma polypeptide；Fc fragment of IgG， low affinityⅢa receptor （CD16a）；free fatty acid receptor 2；interleukin 2 receptor gamma；protein tyrosine phosphatase receptor type C；complement factor B；major histocompatibility complex， classⅡ， DM alpha；major histocompatibility complex， classⅡ， DM beta；major histocompatibility complex， classⅡ， DQ alpha 1；granzyme B. The functions involved the growth and activation of immunologic cell， signal transduction， cell apoptotic， cell factors， receptors， complement， apoptotic， and immunogenicity of tumor cell. Conclusions Down-regulation of a various of immunologic gene expression in colorectal cancer patients with liver metastasis inhibits the function of immunology， and tumor cells escaped the destruction of immunology system results in metastasis.
Objective
To study the differences in gene expression in A549 cells transfected with Forkhead box protein O1(FOXO1),and provide clues to further exploring the mechanism of FOXO1 in acute lung injury.
Methods
After using TNF-α to stimulate A549 cells,the eukaryotic expression vector GV230-FOXO1 was transfected into A549 cells by using lipofectamine transfection reagent.The RNA was isolated and differentially expressed genes were screened with high-throughout DNA microarray.
Results
The eukaryotic expression vector GV230-FOXO1 was successfully constructed and verified.High quality mRNA was isolated and prepared for microarray screening,which passed RNA quality control.The DNA microarray data indicated that 317 genes were up-regulated and 237 genes were down-regulated in A549 cells transfected with FOXO1.The function of these differentially expressed genes involved in many aspects,such as proliferation,apoptosis and differentiation.
Conclusions
Differentially expressed genes in A549 cells transfected with FOXO1 can be successfully screened by using DNA microarray.FOXO1 may influence the progression of the disease by changing the level of cell proliferation,apoptosis and differentiation in acute lung injury.
Objective
To detect and analize the expressions and it’s clinical significance of apoptosis factors in hepatic alveolar echinococcosis tissues by using antibody chip technology.
Methods
The liver tissue specimens (including the edge of lesions and normal liver tissues) of surgical resection of 6 patients with hepatic alveolar echinococcosis in Affiliated Hospital of Qinghai University were collected. The tissue protein was extracted and the level of apoptosis was detected by antibody chip technology. The data were analyzed by AAH-APO-G1 software.
Results
The expression levels of 5 kinds of apoptosis factors (Bad, Fas, IGFBP-3, P21 and XIAP) in the liver tissues of the marginal zone of hepatic alveolar echinococcosis were compared with that of the normal liver tissues, and the difference was statistically significant (P<0.05). The expression levels of Bad, Fas, IGFBP-3 and P21 were up-regulated, and the expression level of XIAP was down regulated.
Conclusions
Apoptosis-related factors play a role in the progression of the hepatic alveolar echinococcosis, there may be contact with the immune escape mechanisms, while promote apoptosis factor and inhibitory apoptosis factor that may exist the function imbalance, so more in-depth exploration the mechanism of apoptosis factors on hepatic alveolar echinococcosis in diagnosis and treatment have important significance.
