Objective The study aimed to explore the main forms of double minute chromosomes(DMs)in the cell cycle of colorectal cancer at different phases,and to clarify the replication and separation phases of DMs.Methods After using serum-free starvation to block the progression of cell cycle in colorectal cancer COLO 320DM cells,Calyculin-A was used to induce interphase cells to prepare karyotype samples through advanced chromatin condensation.COLO 320DM cells were treated with colchicine to ob-tain mitosis(M)phase cells for karyotype analysis.The karyotypes of cells at the early stage of DNA synthesis(G1 phase),the late stage of DNA synthesis(G2 phase),metaphase(M-mid),anaphase(M-late),and telophase(M-ter)of mitosis were observed and photographed under a regular optical microscope,and counted the number of DMs.Results DMs mainly existed in monotypic form at the G1 phase,M-late phase and M-ter phase of cells.In the G2 and M-mid phases of cells,DMs mainly existed in a diploid form.Conclusion Monomeric DMs undergo replication in the S phase and transform from monomers to diploids,while diploid DMs in the M-late phase complete separation and transition from diploids to monomers.

Genetic background can lead to differences in drug effects among different populations when they use the same drug. To delineate the pharmacogenomics and population genetic differences may help to clarify the role of polymorphisms of drug metabolism-related genes in drug effect heterogeneity among different populations. This article has summarized the latest progress on the polymorphisms of drug metabolism-related genes among different populations in China.
China ; Humans ; Pharmaceutical Preparations ; Pharmacogenetics ; Polymorphism, Genetic

Genetic factors play a key role in human athletic ability, and endurance quality and explosive power quality are the important components of athletic ability. In this review, we aimed to reveal the biological genetic mechanism of human athletic ability at the molecular level through summarizing the relationship between genetic variants and human athletic ability, including endurance quality related genetic markers angiotensin converting enzyme (ACE) gene, creatine kinase MM (CKMM) gene and explosive power quality related genetic markers alpha actinin 3 (ACTN3) gene, angiotensinogen (AGT) gene and interleukin6 (IL6) gene. Meanwhile, we also summarized the distribution of allele frequencies among various populations.
Actinin/genetics* ; Athletic Performance ; Gene Frequency ; Genetic Markers ; Genotype ; Humans ; Polymorphism, Genetic

AIK is a novel cationic peptide with potential antitumor activity. In order to construct the AIK expression vector by Gateway technology, and establish an optimal expression and purification method for recombinant AIK, a set of primers containing AttB sites were designed and used to create the AttB-TEV-FLAG-AIR fusion gene by overlapping PCR. The resulting fusion gene was cloned into the donor vector pDONR223 by attB and attP mediated recombination (BP reaction), then, transferred into the destination vector pDESTl 5 by attL and attR mediated recombination (LR reaction). All the cloning was verified by both colony PCR and DNA sequencing. The BL21 F. coli transformed by the GST-AIR expression plasmid was used to express the GST-AIK fusion protein with IPTG induction and the induction conditions were optimized. GST-AIR fusion protein was purified by glutathione magnetic beads, followed by rTEV cleavage to remove GST tag and MTS assay to test the growth inhibition activity of the recombinant AIR on human leukemia HL-60 cells. We found that a high level of soluble expression of GST-AIK protein (more than 30% out of the total bacterial proteins) was achieved upon 0.1 mmol/L ITPG induction for 4 h at 37 °C in the transformed BL21 F. coli with starting OD₆₀₀ at 1.0. Through GST affinity purification and rTEV cleavage, the purity of the resulting recombinant AIK was greater than 95%. And the MTS assays on HL-60 cells confirmed that the recombinant AIK retains an antitumor activity at a level similar to the chemically synthesized AIK. Taken together, we have established a method for expression and purification of recombinant AIK with a potent activity against tumor cells, which will be beneficial for the large-scale production and application of recombinant AIK in the future.
Antimicrobial Cationic Peptides ; biosynthesis ; Antineoplastic Agents ; metabolism ; Escherichia coli ; metabolism ; Genetic Vectors ; HL-60 Cells ; Humans ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Sequence Analysis, DNA

In this paper,a number of valuable experiences focusing on teaching idea,cultivation of high-quality teachers and deepening of teaching content and method reform were summarized through exploring the construction and practice of the national excellent course-medical genetics. Aim of the ex-ploration is provide references for the construction and application of national excellent course and its sus-tainable development.

Objective To investigate the effect of arsenic trioxide on human coronary smooth muscle cells(HCSMCs). Method After the subculture of HCASMCs, cells were divided into 4 groups: control group and three arsenic trioxide (4.0 μmol/L) groups as the arsenic agent co-cultured with cells for 12,24 and 48 h. Flow cytometry and TUNEL were used for counting the ntmnber of cell apoptosis. RT-PCR was used to detect the expression of E2F-1 mRNA and Western blot was used to measure the levels of Bcl-2 ond Bax. Results Arsenic trioxide inhibited the proliferation of HCASMCs. When arsenic trioxide was 4.0 μmol/L, the living cells were ( 8.44 ±0.10) × 105/mL,signifiantly than that of control group (16.44 ± 1.34) × 105/mL, P ＜ 0.05. This inhibition of proliferation cycle was produced by affecting S and G2-M phase, which did not appear in the control group. Apoptotic cells increased from 16.0% ± 3.1% to 38.7% ± 2.7% (P ＜ 0.05) detected by TUNEL. The arsenic agnet decreased the Bcl-2 level and increased Bax. The expression of E2F-1 mRNA was decreased in 4.0 μmol/L arsenic trioxide group. Conclusions Arsenic trioxide exerts the apoptotic effect on human coronary smooth muscle cells.

Objective To investigate the apoptotic effects of arsenic trioxide on E2F-1 and EMAP-Ⅱ of hu-man coronary artery smooth muscle cells (HCASMCs). Method HCASMCs proliferated after culturing cells.Cells were divided into 4 groups: control group and arsenic trioxide (4.0μmol/L) 12 h group, 24 h group and 48 h group. Flow cytometry was used for counting number of apoptotic cells, RT-PCR was used for detecting the ex-pression of E2F-1 mRNA and Western blot was employed to get the level of EMAP-Ⅱ. Results Arsenic trioxide inhibited the proliferation of HCASMCs (living cells in 4.0 μmol/L arsenic trioxide were (8.44±0.10)×10~5/mL vs. control (16.44±1.34)×10~5/m (P <0.05), and the 4.0μmol/L arsenic trioxide inhibited proliferation cy-cle via affecting S and G2M phases, while those were not found in control group. E2F-1 mRNA expression was de-creased in 4.0 μmol/L arsenic trioxide group. Western blot test showed EMAP- Ⅱ level was also decreased in 4.0 μmol/L arsenic trioxide group. Conclusions Arsenic trioxide has apoptotic effect on smooth muscle cells of hu-man coronary artery and decrease the expression of E2F-1 mRNA and the level of EMAP- Ⅱ.

BACKGROUND: Limb allograft is a sort of composite tissues allotrans plantation(CTA), some researches showed that the apoptosis of target cell is one of the main mechanism of the dysfunction of allograft.OBJECTIVE: To investigate the characteristic of cell apoptosis in acute rejection of limb allograft in rats based on limb allograft model.DESIGN: A randomized controlled trial using the experimental animals as the objects.SETTING: Experimental animal center Laboratory of a hospital of a medical university MATERIALS: The experiment was done in the Experimental Animal Central Laboratory of the First Affiliated Hospital of Harbin Medical University from November 2003 to May 2004. Totally 56 healthy and male SD rats and 28 Wister rats were involved with body mass of 200 to 250 g. The rats were provided by the Experimental Animal Center of the First Affiliated Hospital of Harbin Medical University. They were randomly divided into two groups:transplantation group with 28 Wistar rats and 28 SD rats and control group with 28 SD rats.INTERVENTIONS: The transplantation group of SD rats underwent limb allotransplantation from allogenetic Wistar. The control group of SD rats underwent limb replantation. The expression of acute rejected in limb allografts was observed. The limb grafts were harvested atday 1, 3, 5 or 7 after transplantation. Histopathological rejection grade of each tissue rejection was performed with hematoxylin-eosin staining. Apoptotic cells were detected by TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling) and apoptotic index(AI) was calculated.MAIN OUTCOME MEASURES: Primary results: ① istopathological grade of acute rejection in limb allografts ② The relationship between apoptosis and acute rejection in limb allograft in rats; Secondary results:General condition of rats in each group.RESULTS: The limb grafts showed edema and erythema and the skin became red at day(3.43 ±0.79) after transplantation. The average survival time was(7.42 ± 1.72) days. The acute rejection in skin was the strongest. On the day 3, 5 and 7 after operation, the histopathological rejection grades of skins in the transplantation group were(1.14±0.38) ,(2.28 ±0.48) and(2.86 ±0.38) grades respectively. They were significantly different from that of muscle and nerve( P ＜ 0.05 ) . The apoptotic cells in allografts were mainly infiltrating lymphocytes in subcutaneous tissues and then the muscle cells. All was positively correlated with acute rejection grade in limb allograft .CONCLUSION: Apoptosis was involved in acute rejection of limb allograft in rat. The apoptotic index can be used as a quantitative index to estimate the injury of grafts.

OBJECTIVETo search the candidate gene in the development and metastasis of lung adenocarcinoma and shed light on the possible molecular mechanism of the development of lung carcinoma.

METHODSUsing methods of cell culture, reverse transcription-PCR, RH gene mapping and RNA in situ hybridization.

RESULTSThe cDNA fragment named OPB7-1 was mapped at 1p31-1p34 by RH gene mapping method. The fragment sequences obtained from lung cDNA library of normal person and cell line of AGZY83-a were similar in length but showed individual base difference. For OPB7-1, there is a low homogeneity to known gene by analysis in GenBank, but 3 contigs homologous to OPB7-1 were located at chromosome 1(1p31-1p34). Different degrees of expression were noted in tumor tissues from 24 cases of lung carcinoma, however no significant expression was found in their corresponding normal tissues. And high expression was found in the lung tissues of cases with lymph node metastasis.

CONCLUSIONOPB7-1 may be a novel gene. It may be a tumor related gene in occurrence and metastasis of lung carcinoma.

Adenocarcinoma ; genetics ; Animals ; Chromosome Mapping ; Cloning, Molecular ; DNA, Complementary ; genetics ; Gene Expression Regulation, Neoplastic ; Genetic Predisposition to Disease ; genetics ; Humans ; In Situ Hybridization ; Lung Neoplasms ; genetics ; pathology ; RNA, Neoplasm ; genetics ; metabolism ; Radiation Hybrid Mapping ; Rats ; Tumor Cells, Cultured

OBJECTIVETo investigate common chromosomal changes and the LOH frequency of microsatellite loci in primary gastric cancer samples in order to locate the deleted regions in which human gastric cancer related genes might exist.

METHODSComparative genomic hybridization (CGH) was used to define global chromosomal aberrations in 43 primary gastric tumors. Based on the results of CGH, analysis of loss of heterozygosity (LOH) was performed in chromosome 19 in which the loss was first discovered in the gastric cancers. The PCR-based approach was used to investigate 22 loci, which are spaced at 1.1 - 10.9 cM intervals throughout chromosome 19. The amplified PCR fragments were subjected to electrophoresis in PAGE gel and analyzed with Genescan trade mark and Genotyper trade mark.

RESULTSCGH analysis revealed gains in chromosome 3p (8/43), 8q (8/43), 20 [20 (9/43), 20p (7/43), 20q (4/43)], 12q (16/43), 13q (12/43) and losses in 19 [19 (15/43)], 7 [17 (8/43), 17p (10/43)], 16 (10/43) and 1p (11/43). Among the 43 evaluated samples, the most frequent LOH was detected at locus D19S571 (27.81%).

CONCLUSIONSThe tumorigenesis of gastric cancer includes several chromosomal changes. The aberration of chromosome 19 was the first common change founded in gastric cancer. The region near the D19S571 might harbor potential genes related to the tumorigenesis of gastric cancer.

Chromosomes, Human, Pair 19 ; Humans ; Loss of Heterozygosity ; Nucleic Acid Hybridization ; Peutz-Jeghers Syndrome ; genetics ; Stomach Neoplasms ; genetics