Advance in research on genetic polymorphisms associated with athletic ability / 中华医学遗传学杂志

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.

Progress of research on polymorphisms of common drug metabolism related genes among various populations in China / 中华医学遗传学杂志

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.

Expression, purification of recombinant cationic peptide AIK in Escherichia coli and its antitumor activity / 生物工程学报

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.

Establishment and practice of the national excellent courses of medical genetics / 中华医学教育探索杂志

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.

Effects of arsenic trioxide on E2F- 1 and EMAP-Ⅱ of smooth muscle cells of human coronary artery in vitro / 中华急诊医学杂志

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.

Apoptostic effect of arsenic trioxide on human coronary smooth muscle cells in vitro / 中华急诊医学杂志

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.

Apoptosis in the process of limb allograft in rats / 中国组织工程研究

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.

Analysis of comparative genomic hybridization and loss of heterozygosity in 43 primary gastric carcinomas / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Comparative 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.</p><p><b>RESULTS</b>CGH 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%).</p><p><b>CONCLUSIONS</b>The 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.</p>

Cloning and mapping analysis of cDNA fragment OPB7-1 gene in human lung adenocarcinoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Using methods of cell culture, reverse transcription-PCR, RH gene mapping and RNA in situ hybridization.</p><p><b>RESULTS</b>The 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.</p><p><b>CONCLUSION</b>OPB7-1 may be a novel gene. It may be a tumor related gene in occurrence and metastasis of lung carcinoma.</p>

The effect of RAB5A gene over-expression on invasive and metastatic capabilities of human lung adenocarcinoma cell lines / 中国肺癌杂志

<p><b>BACKGROUND</b>Over-expression of RAB5A gene has been proved to be associated with neoplasia metastasis. This study is to explore the effect of RAB5A gene on invasion and metastasis of human lung adenocarcinoma cell lines.</p><p><b>METHODS</b>Constituted basement membrane invasion technique, adhesion capability of tumor cell assay, the chemotactic migration of tumor cells assay, and gelatinases SDS-PAGE analysis method were used to detect the changes of invasive and metastatic capability of Anip973 (with high metastatic capability) and its parent AGZY83-a cell lines (with low metastatic capability).</p><p><b>RESULTS</b>After AGZY83-a cells were transfected by PcDNA3.1-RAB5A plasmid, its invasion was significantly increased (t=24.36, P < 0.000 5); adhesion capability of cell was promoted (P < 0.05); the chemotactic migration of cells was higher than that of the parent lines (t=14.18, P < 0.000 5); and the activity of gelatinases secreted from transfected AGZY83-a was enhanced. The invasion of the transfected Anip973 cells with PcDNA3-AntiRAB5A was lower (t= 16.510 4, P < 0.002 5); adhesion capability of cell was decreased (P < 0.05); the chemotactic migration of cells was lower than that of the parent lines (t=6.062, P < 0.005); and the activity of gelatinases was obviously decreased.</p><p><b>CONCLUSIONS</b>This study in vitro indicates that over-expression of RAB5A genes plays an important role in tumor invasion and metastatic phenotype formation of human lung adenocarcinoma cells; and antisense RNA can interrupt the translation of RAB5A gene.</p>

Genescan analysis of non-small cell lung cancer in the long arm of chromosome 6 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate if there are microsatellite loci in the long arm of chromosome 6 that have close relationship with non-small cell lung cancer.</p><p><b>METHODS</b>Multiple PCR approach was used to analyze the 18 loci in the long arm of chromosome 6. The PCR products were analyzed in PAGE, and then the electrophoresis maps were analyzed with Gene Scan(TM) and Genotyper(TM).</p><p><b>RESULTS</b>There were different frequencies of loss of heterozygosity (LOH) in different loci (varying from 3.85% to 38.45%). The total frequency of LOH in 41 gastric cancers was 58.5%(24/41). Eight loci with the LOH frequency higher than 20% were mainly located in 2 regions: 6q24 and 6q27. The accurate location is 6q24-6q25.3 [D6S1699(35%), D6S409(23.33%), D6S441(33.33%)] and 6q26-27 [D6S1550(38.45%), D6S264(20%), D6S1585(25%), D6S446(33.33%), D6S281(30.77%)].</p><p><b>CONCLUSION</b>There may be tumor suppressor genes located in the region of 6q24 and 6q27, which have close relationship with non-small cell lung cancer.</p>

Effect on radiosensitivity of human lung adenocarcinoma cells transfected with wild-type p16 gene / 中国癌症杂志

0.05); The result of FCM indicates that the cells transfected with p16 changed in cell cycle distribution ; proportion of G 2 M phase was increased, while that of S phase was decreased.Conclusions:Introduction of p16 gene into low expression human lung adenocarcinoma cell line Anip973 alters the radiation effect in a way which perhaps was correlated with the inhibition of sublethal damage repair and the cell cycle distribution change, but not in the high expression cell line AGZY83a.

RAT P53GENE MUTATIONS IN THE COLORECTAL TUMORS INDUCED BY N-METHYL-N-NITROSOUREA / 解剖学报

Objective We have examined tumor suppressor gene p53 from N- methyl - N- nitrosourea (MNU ) induced the col - orectal tumors in rats. Methods PCR, SSCP and DNA sequence analysis. Results In colorectal tumors induced by MNU, the mobil- ity shifts of p53 gene mutation were observed, p53 gene mutation was a deletion at third position C of codon 244 in exon7. Conclu- sion The present study suggested that MNU induces rat p53 mutations in the colorectal tumors.