Epidermal growth factor receptor (EGFR) transcript level in breast cancer tissues

Background: Epidermal Growth Factor Receptor (EGFR) is a transmembrane cell-surface glycoprotein with intrinsic tyrosine kinase activity. EGFR has been shown to stimulate cell proliferation and to enhance the migration and invasiveness of breast cancer. EGFR is expressed in epidermal cell lines and have been implicated in the pathogenesis of many different types of cancer. Objective: To evaluate the level of EGFR transcript in breast cancer and normal tissues; comparison the EGFR transcript level at different development stages and cancer cell types. Subject and methods: Total RNA from 62 tissue samples including 47 breast cancer and 15 normal tissues were extracted; cDNA synthesis by reverse transcript polymerase chain reaction, EGFR transcript level were determined using semi-quantitative RT-PCR. Result and conclusions: EGFR transcript level was highly expressed in breast cancer tissues compared to the normal tissues. Especially, its expression was related to the different status and cancer cell types of breast cancer. There was a difference of EGFR transcript level between histological pathology\u2019s forms of breast cancer in the same stage.

Detection of mutant dystrophin gene carrier using quantitative Polymerase Chain Reaction

Background: Deletion and duplication mutations of dystrophin gene make up from 70 to 75% of patients with Duchenne Muscular Dystrophy (DMD). Two thirds of children with DMD inherited from the heterozygous mothers the mutated gene which is located on one of the sex chromosomes. Objective: To detect the asymptomatic carriers of dystrophin gene mutation using molecular techniques. Subject and methods: 3 DMD patients and their 9 relatives. Using techniques: DNA extraction and quantitative Polymerase Chain Reaction (PCR). Results: Successfully detected 4 heterozygous individuals from 9 female members of three different families that have already confirmed DMD patients. Conclusion: This method could lead to a new way of prenatal diagnosis of DMD as well as other genetic disorders that are caused by deletion or duplication mutation.

Cloning of the gene encoding human recombinant blood \u2013 coagulation factor WIII

Background: Hemophilia A is a genetic bleeding disorder that results from a deficiency in factor VIII. The prevalence of Hemophilia in Vietnam is rather high (2/34830 people) and Vietnam has high usage demand for factor VIII in the treatment and prevention of the disease. Therefore, it is necessary to study and produce recombinant blood \u2013 coagulation factor WIII. Objective: To clone successfully A1A2 and A3C2 gen fragment encoding factor VIII. Subject and Method: Amplify A1A2 and A3C2 gene fragments by PCR from human cDNA. PCR products were ligated into cloning vector pQE \u2013 30UA. Recombinant plasmids were transformed into E.coli DH5 alpha host strain. Inserted A1A2 and A3C2 gene fragments were checked by PCR and restriction enzymes. Result: Successfully amplifying functional gene fragments encoding factor VIII using specific primers. Conclusion: Obtaining pQE \u2013 30UA vector carrying A1A2 and A3C2 fragments encoding factor VIII. This is the premise result for the next studies on synthesis of recombinant factor WIII and application of genetic therapy.

Expression of Heparansulfate Interacting Protein (HIP) in benign prostatic hyperplasia,prostate intraepithelial neoplasia and prostate cancer

Background: Heparansulfate Interacting Protein (HIP) is up-regulated in various human cancer cell lines at both transcript and protein levels. HIP expression is related to the differentiation status and cancer development. Objectives: To determine HIP in benign prostatic hyperplasia, prostatic intraepithelial neoplasia and prostate cancer tissues. Materials and method: Western blot method was used to determine HIP expression in 3 different types of prostate tissue, including 11 prostate cancer samples, 2 benign prostatic hyperplasia samples and 11 prostatic intraepithelial neoplasia samples. Results. HIP was particularly up-regulated in prostate cancer and prostatic intraepithelial neoplasia, indicating that up-regulation of HIP expression may be an early event in tumorgenesis. Conclusion: The expression of HIP was different between cancer, prostatic intraepithelial neoplasia tissue and benign prostatic hyperplasia. HIP may serve as a prognostic marker for prostate carcinoma.

Up-regulate Heparansulfate Interacting Protein (HIP) transcript in breast cancer tissues

Background: Heparansulfate Interacting Protein (HIP) is a protein that belongs to a novel class of heparin and heparansulfate binding protein. It plays an important role in extracellular matrix structure and function, cell-cell and cell-extracellular matrix adhesion, growth and differentiation. HIP was shown to be expressed in normal epithelia and epithelial cell lines at both mRNA and protein levels. Especially, HIP was found to be up-regulated in some cancer cell lines and related to different status and metastasis.\r\n', u'Objectives: To determinate HIP transcript level of mRNA in breast cancer tissues in comparison with normal tissues; to compare HIP transcript level at different cancer stages and cancer cell types. \r\n', u'Subjects and method: Total RNA was isolated from 62 tissue samples (47 breast cancer and 15 normal tissues); cDNA synthesis by reverse transcript \u2013polymerase chain reaction (RT \u2013 PCR); determination of HIP transcript using semi-quantitative RT \u2013 PCR. \r\n', u'Results: HIP transcript was particularly up \u2013 regulated in breast cancer tissues compared to normal tissues, especially this up-regulated in cancer tissues at different stages of development and cancer cell types. \r\n', u'Conclusion: These results show that the HIP transcript level was different between breast cancer and normal tissues and its expression was related to different status and metastasis in human cancer cell lines. HIP may be used as a prognostic marker for breast cancer.\r\n', u'

A nonsense mutation effects mRNA splicing process of dystrophin gene

Background: Production of semi-functional dystrophin protein from the dystrophin gene encoded with a premature stop codon has been shown to modify the severe phenotype of Duchenne Muscular Dystrophy (DMD). The mutation of the dystrophin gene affects the process of complete mRNA and is important in gene therapy. Objective: To analyze the mutation of dystrophin gene in DMD cases. Subjects and methods: A patient with diagnosed with DMD when he was 2 years old, and at age 9, he was completely disabled and had to use a wheelchair. DNA and total RNA were extracted from fresh peripheral blood; cDNA was synthesized by transcript polymerase chain reaction (RT - PCR). PCR, nested PCR or sequence methods were used to determine the mutation of the dystrophin gene. Results: A nonsensical mutation (E638) due to a single nucleotide change in exon 17 of the dystrophin gene (GAA2047TAA) was identified. This mutation affects mRNA splicing process and induces complete exon 17 skipping. Conclusion: Patients, who had E638X mutation with exon 17 deletion in the dystrophin gene, had clinical symptoms of Becker Muscular Dystrophy (BMD). This discovery as a potential target for therapeutic strategies for DMD, to change the severe phenotype of DMD to a milder phenotype (BMD), in order to improve clinical conditions for the patients.

The differences of transcript level of Heparan-sulfate Interacting Protein (HIP) in benign prostatic hyperplasia, prostatic intraepithelial neoplasia and prostate cancer tissues

Background: Heparan-sufate interacting protetin (HIP) has been known to be up-regulated and expressed in various human cancer cell lines at both transcript and protein levels. HIP\u2019s expression was related to the differentiation status and cancer development. Objective: Using a semi-quantitative PCR method to determine HIP transcript levels in benign prostatic hyperplasia (BPH), prostatic intraepithelial neoplasia (PIN) and prostate cancer tissues. Subjects and methods: 30 samples of BPH, 12 samples of high-grade PIN, and 40 samples of prostate cancer were collected from patients at Viet Duc Hospital and Friendship Hospital. Total RNA was extracted from BPH, PIN and prostate cancer tissues; cDNA synthesis by reverse transcript - polymerase chain reaction (RT - PCR); HIP transcript determination using semi-quantitative PCR. Results: There was significant difference in HIP transcript levels. HIP transcript was very highly up-regulated in the prostate cancer tissues. The up-regulation of HIP transcript was lower in PIN, and lowest in BPH. HIP transcript levels in benign samples were 1/2 and 2/3 compared with cancer and PIN samples, respectively (P< 0.05). These indicated that up-regulation of HIP transcript may be an early event in tumorigenesis. Conclusions: Levels of HIP transcript were different between tissues of prostate cancer, PIN, and BPH. HIP may be a marker for pre-cancer of the prostate.

The determination heparansulphat interacing protein (HIP) transcript in cancer tissues using semiquantitative PCR and quantitative PCR methods

Background: Semiquantitative PCR and quantitative PCR are accurate and simple methods. They are commonly used to determine amplified gene levels in PCR reaction. Objective: Using semiquantitative PCR and quantitative PCR methods to determine HIP transcript levels in cancer and normal tissue; to evaluate sensibility of two methods. Subject and methods: 30 cancer patients were diagnosed based on clinical, para-clinical (ultrasound, biochemistry, histopathology) at K hospital in Hanoi. 15 benign tissue samples are used for control.mRNA was extracted from cancer and normal tissues, cDNA was synthesized by reverse transcript-polymerase chain reaction (RT - PCR); HIP transcript was determined using semiquantitative PCR and quantitative PCR methods. Results: Both methods showed the same results: HIP transcript was increase in cancer tissues but very low in normal tissues. This showed that HIP was linked closely with the development of cancer tissue. Conclusions: Levels of HIP transcript was different between cancer tissue and the normal control. Semi- quantitative PCR and quantitative PCR are useful methods to determine HIP transcript for cancer diagnosis. \r\n', u'\r\n', u'\r\n', u'

Spoligotyping technique: improvement and application in classification Mycobacterium tuberculosis

Background: There are many methods used in epidemiological studies of tuberculosis (TB) bacteria but Spoligotyping method is widely used with high accuracy, simple procedure, and carried out on strains containing a little of IS6110 segment \r\n', u'Objectives: To improve Spoligotyping technique and apply this technique to classify Mycobacterium tuberculosis\r\n', u'Subjects and method: Subjects and methods: The study included 12 medical waste samples collected at Thai Binh Hospital of Tuberculosis and 19 samples obtained from the The Hanoi Institute for Tuberculosis and Lung Diseases. Spoligo model of 31 samples were analyzed based on Spotclust and SpoIDB4 database and divided up into family and subfamily.\r\n', u'Results: Spoligotyping technique has good results with the PCR product amplified 40 cycles and presented the film in 18 hours. Obtained results in 31 medical waste samples belong to 4 families: Beijing, EAI, T1 and H3-LAM9. EAI and Beijing are dominant families with 45.16% and 38.7%), respectively. T1 and H3-LAM9 are 12.9% and 3.22%, respectively. The number of samples in the study is little but the obtained rate of different spoligo models of strains are quite diversified (41.9%)\r\n', u'Conclusion: This result is relatively appropriate with previous studies on the distribution of EAI and Beijing families in Vietnam and the world. Spligotyping technique distinguished samples belonging to Beijing or non-Beijing families, which support for clinical treatment and development of new vaccines.\r\n', u'\r\n', u'\r\n', u'