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Objective: To investigate the effect of Dahuangzhechong pill on the gene expression spectra of preventing arterial thrombosis, and reveal its mechanism on molecule level., Methods: Mononuclear cell and blood platelet of the arterial thrombosis patients were separated before and after treatment by Dahuangzhechong pill. Their RNA was extracted respectively and the genes expressions were detected using gene array containing 14,000 gene., Results: 44 genes up-expressed and 299 genes down-expressed in blood platelet, 252 genes expression increased and 299 genes expression decreased in mononuclear cell genes after treated with Dahuangzhechong pill. The cluster analysis showed that the genes contained ion channel and transport protein, apoptosis related protein, DNA synthesis, repair and transcription factor, cell receptor, cell signal and transducin, and protein translation and synthesis, etc., Conclusion: Dahuangzhechong pill may prevent arterial thrombosis through genes containing ion channel and transport protein, apoptosis related protein, DNA synthesis, repair and transcription factor, cell receptor, cell signal and transducin, and protein translation and synthesis, etc.

Objective: To establish and evaluate a rabbit model of arterial thrombosis by modified thread-drawing., Methods: Fifty-three rabbits were randomly divided into 6 groups: a normal group, a ligating group,a collagen encapsulated thread-drawing group,a aspirin group,a clopidogrel group, and an aspirin clopidogral group. The endovascular pathological changes in the rabbits were observed, and D-fructose-1,6-diphosphate trisodium salt octahydrate (FDP), D-Dimer and tissue factor (TF) were detected with enzyme linked immunosorbent assay., Results: In the thread-drawing group, thrombus was obvious, and the endovascular elastic membrane was injured seriously compared with the ligating group. After being treated with aspirin and clopidogrel, most arterial thrombus was softened, dissolved and absorbed. Compared with that in the modified thread-drawing group,wet and dry weight of thrombus increased,and the level of D-Dimer, FDP and TF also increased in the modified thread-drawing group (P<0.01). After being treated by aspirin and/or clopidogrel, the wet and dry weight of thrombus and the level of D-Dimer, FDP and TF decreased compared with the control (P<0.01). Aspirin plus clopidogral could obviously reduce the wet and dry weight of thrombus, and reduce the level of D-Dimer and FDP (P<0.01). Aspirin plus clopidogral could obviously inhibit the formation of TF compared with aspirin (P<0.05)., Conclusion: Arterial thrombosis model by collagen encapsulated thread-drawing which is visible, repeatable and effective is better than thread-drawing. It is suitable for screening anti-thrombosis drugs and evaluating their effect.

Objective: To explore the molecular mechanism of Glanzmann thrombasthenia (GT)., Methods: All 45 exons of alphaIIb and beta3 subunit genes as well as their splicing sites were amplified by polymerase chain reaction(PCR) with 40 primer pairs, and then the PCR products were used to screen the gene mutation by single strand conformation polymorphism-polyacrylamide gel electrophoresis (SSCP-PAGE). The mutation was further confirmed by direct DNA sequencing., Results: A DNA band alterated migration was detected after SSCP-PAGE. DNA sequencing showed that a base deletion within the band at the site of 540 in GPIIb gene(540A) was found., Conclusion: The frame-shift mutation caused by the deletion of 540A in GPIIb gene is a novel mutation which is a genetic defect in patients with GT.

Aim: To establish the two-dimensional gel electrophoresis (2-DE) profiles of indomethacin (IN)-treated human colon cancer cell line HCT116, and to provide a new way to study its anti-tumor molecular mechanism through analyzing a variety of protein maps., Methods: Two-DE profiles of HCT116 were established in IN-treated and untreated groups. Total proteins were separated by immobilized pH gradient-based 2-DE. The gels were stained by silver, scanned by ImageScanner, and analyzed with Image Master software., Results: Clear background, well-resolved and reproducible 2-DE patterns of HCT116 cells were acquired in IN-treated and untreated group. The average deviation of spot position was 0.896+/-0.177 mm in IEF direction and 1.106+/-0.289 mm in SDS-PAGE direction respectively. In IN-treated group, 1169+/-36 spots were detected and 1061+/-32 spots were matched, the average matching rate was 90.6% in three gels. In untreated group, 1256+/-50 spots were detected and 1168+/-46 spots were matched, the average matching rate was 93.0% in three gels. Forty-five differential protein spots were displayed between IN-treated and untreated groups. Of which, 34 protein spots decreased and 9 showed higher expression in IN-treated group, and only two protein spots showed an expression in untreated cells., Conclusion: Two-DE profiles of IN-treated and untreated HCT116 cells were established. Apparent 45 different protein spots were detected in IN-treated and untreated HCT116 cells. The analysis on differential protein spots may serve as a new way to study the molecule mechanism of IN-treated colon cancer.

Objective: To find out the differentially expressed genes in the hippocampus of the rats with genetic epilepsy so as to lay a foundation for exploring the pathogenesis of epilepsy by means of cDNA array technology., Methods: Gene expression patterns in the hippocampus of the genetic epilepsy-prone P77PMC rats and normal Wistar rats were established using the alpha-32P-labeled cDNA probes hybridized with the Atlas Rat cDNA Expression Array, and then were analyzed by an image analysis instrument to get the differentially expressed genes., Results: Fifteen genes were found having differential expression patterns in hippocampus between the P77PMC rats and the Wistar rats, while there may be many other differentially expressed genes left undiscovered due to having no appropriate image analysis software. And among the fifteen genes, the expression levels of twelve genes in the P77PMC rats were higher than those in the Wistar rats, while the expression levels of the other three genes were lower. The results of reverse transcription-polymerase chain reaction(RT-PCR) have demonstrated the reliability of cDNA arrays method., Conclusion: cDNA array is a powerful tool for identifying differential expression genes of epilepsy on large scales. There are several differentially expressed genes in hippocampus of the P77PMC rats and the Wistar rats. All these identified genes could play potentially important roles in the pathogenesis of epilepsy.

Objective: To investigate the inhibitory mechanism of Yiqijiedu granule on the implanted-tumor growth of nasopharyngeal carcinoma in nude mice., Methods: Twenty nude mice were injected nasopharyngeal carcinoma cells (HNE1), with 5 x 10(6) cells for a nude mouse. Implanted-tumors grew for 20 d, whose volume reached 1 cm x 1 cm x 1 cm. These nude mice were divided into 2 groups: Yiqijiedu group and control group. The Yiqijiedu group was given Yiqijiedu granule, and the control was given normal saline for 30 d, and then were killed. The volume and weight of implanted-tumors were measured. A 100-mg tissue from implanted-tumors was used to extract total protein by current methods, in which the proteins were separated by two-dimension electrophoresis and stained by silver, and protein profiles of implanted-tumors were obtained. Different proteins in the profiles were analyzed by ImageMaster 2D Elite 4.01. Nineteen different proteins, in which 4 expressed in the Yiqijiedu group, 4 expressed in the control, and the other 11 were differently expressed at 5 folds, were identified by MALDI-TOF-MS., Results: The Yiqijiedu granule could inhibit the growth of implanted-tumors. The volume and weight of implanted-tumors in the Yiqijiedu group were (0.207 +/- 0.023) cm3 and (0.132 +/- 0.021) g respectively, and that of the control was (1.342 +/- 0.298) cm3 and (1.017 +/- 0.015) g ( P < 0.05). The inhibitory rate was 84.58%. The analyses of two-dimension electrophoresis and ImageMaster 2D Elite 4.01 showed that there were 567 +/- 49 protein dots in the Yiqijiedu group and 679 +/- 59 in the control. We found 243 proteins were dys-regulated, of which 112 proteins were observed, up-regulated and the other 131 proteins were down-regulated. MALDI-TOF-MS and Database analysis showed that the high expression proteins were HKR2 protein, 10 Phosphoribosyl pyrophosphate synthetase, TNFR superfamily member, and Apoptosis regulator. The lower expression proteins were Fibulin-3, Zinc finger protein 266, Carboxy terminus of HSP70-interacting protein, et al., Conclusion: Yiqijiedu granule could inhibit the growth of nasopharyngeal carcinoma, which may be associated with those proteins regulated by itself.

Background & Objective: Lung cancer is a kind of disease with high incidence and mortality, however its molecular mechanism is not clear yet. This study was designed to investigate gene expression differences among lung cancer tissues, lung paracancerous tissues,matched peripheral normal lung tissues and the metastases of lymph nodes, and to seek the relatively high expressed genes in lung carcinoma tissues, providing possible theoretical basis for early diagnosis and treatment of pulmonary carcinoma., Methods: Fresh lung cancer tissue, lung paracancerous tissue, matched normal lung tissue and metastases of lymph nodes were deep-frozen in liquid nitrogen; their total RNA were extracted for reversed transcription cDNA probes, which were labeled and subsequently used to hybridize with cDNA microarray with 588 genes. Different gene expression profiles were obtained by analyzing the integrated density (ID) of spot images on the X-ray., Results: In lung cancer tissues, 40 genes were detected to be differentially expressed, 36 of which such as early growth response protein 1 (EGR1), secreted apoptosis related protein 1 (SARP1) were upregulated while the others such as myeloid cell leukemia protein 1 (MCL1) were downregulated. The upregulated genes were mainly oncogene/suppressor gene, cell cycle regulatory gene, growth factors and apoptosis-related genes. In lung paracancerous tissues, 33 genes had different expression, 20 of which such as matrix metalloproteinase-9 (MMP-9) were upregulated, and the other 13 such as MCL1,endothelin 2 (ET2) were downregulated. In metastases of lymph nodes, there were 21 genes found to be differently expressed, 15 of which such as CD40 receptor-associated factor 1 (CRAF1) were downregulated while the rest were upregulated, and the upregulated genes (6 in 15) were mainly the genes associated with adhesion molecules, matrix metalloproteinases and collagen., Conclusion: EGR1, SARP1, NDKA, etc. may be the key genes in pulmonary carcinogenesis course. At the same time, MMP9, thrombospondin 2 (TSP2), etc.may play an important role in pulmonary cancer metastasis and infiltration.

Objective: To investigate the effect of Coptis Chinensis compound on the gene expression of transplanted tumor in nasopharyngeal carcinoma cell line of CNE1., Methods: The cells were injected into Balb/C nude mice, and after the transplanted tumor was established, the nude mice with tumors were treated with Coptis chinensis compound. The nude mice were then killed, tumor tissues were isolated and the total RNA was extracted. Hybridization with a cDNA microarray was conducted using a probe of fluorescent-labeled cDNA mixture that was reversely transcribed from the RNA. The gene expression level was analyzed after scanning the fluorescent intensity by a laser scanner, and further confirmed by RT-PCR., Results: After using Coptis Chinensis compound for 30 days, the size of transplanted tumors were remarkably reduced, and the inhibition rate of tumor growth was 29.5%. In the meantime, the expression of 147 genes in the tissue of implanted tumor treated with the compound altered: the expression of 102 genes was down-regulated, and that of 45 genes was up-regulated. Furthermore, the differential expression of 3 genes was verified by RT-PCR, MAD3 and H731 genes were up-regulated, and that of CHK1 was down-regulated., Conclusion: Coptis Chinensis compound can affect the gene expression in the tissue of implanted tumor, which implies that the inhibition of Coptis Chinensis compound on the growth of the implanted tumor with CNE1 cells in nude mice might be associated with the control of gene expression.

Background & Objective: Previous studies showed that JNK signaling pathway activated by LMP1 plays an important role in carcinogenesis of nasopharyngeal carcinoma (NPC). JNK interacting protein (JIP) can inhibit JNK signaling pathway in NPC cell. This study was designed to elucidate the effect of JIP on the proliferation and apoptosis of NPC cells., Methods: After treatment with JIP at different concentrations and time points, the number of proliferating cells were determined by MTT assay; the ability of proliferation of NPC cells was measured by the rate of cloning formation; cell cycle and the apoptotic rate of NPC cells was assayed by flow cytometry., Results: 1. MTT assay showed that cell proliferation was significantly inhibited by JIP in a dose- and time-dependent manner. After treatment with JIP for 24, 48, and 72 hours, the rate of survival cells were 77.8%, 59.2%, and 61.8%, respectively. 2. The number and volume of colonies were decreased after transfection with JIP. 3. The number of cells in S phase was decreased from 25.87% to 19.96%, and the number of cells in G0/G1 phase was elevated from 66.24% to 71.89% after treatment with JIP. 4. In contrast to the control group, the 24 hours apoptotic rate was elevated from 1.25% to 8.25% (about 6.6 times); the 48 hours apoptotic rate was elevated from 1.04% to 31.45% (about 30 times)., Conclusions: The results demonstrated that JIP inhibit the growth of nasopharyngeal carcinoma through arresting the cell cycle at G1/S checkpoint and triggering the apoptosis of cells. Data suggest that JIP is a potent molecular drug for the treatment of the patients with nasopharyngeal carcinoma.