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Objective To investigate the expression and biological function of miRNA-449 a in lung cancer . Methods A case-control study was conducted in 58 patients diagnosed with lung cancer ( carcinoma and adeno-carcinoma) and normal tissue closely adjacent to tumor.MiRNA-449a simulation was designed and synthesized, was dissolved into two different concentrations as 10 and 20 mg/mL.The expression of miRNA-449a in lung cancer tissues and matched normal tissues were detected by Real time PCR .The expression of luciferase gene was detected by chemiluminescence technique.MiRNA-449a mimics on cell apoptosis was evaluated by MTT assay . Results The mean tissues expression levels of miRNA-449 a in squamous carcinoma group and adenocarcinoma group were 1.48 ±1.63 and 1.52 ±1.54 respectively, and were significantly lower than in control group (2.74 ± 1.55 ) ( P<0.01 ) .The average intensity of fluorescent protein in 10 mg/mL group and 20 mg/mL group were 2 115 ±168 and 1 352 ±159 respectively , and were significantly lower than that in control group ( 4 975 ±115 ) ( P<0.01 ) .Conclusions MiRNA-449 a was down-regulated expression in lung cancer and induced apoptosis .
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Objective To understand the detection situation of Mycoplasma pneumoniae (MP)and Chtamydia trachomatis (CT) in children with respiratory tract infection.Methods MP and CT were simultaneously measured by fluorogenic quantitative PCR in 1393 children patients with respiratory tract infection.Results The total detection rates of MP and CT were 30.4%,in which MP was 21 .8%,CT was 9.3% and MP complicating CT was 0.65%.In the season distribution,the detection rates of MP and CT were highest in summer.In the age distribution,the detection rate of MP in the over 3-year-old group was higher than that in the other age groups,the differences had statistical significance(P <0.05).The detection rate of CT in the less than 1-month-old group was higher than that in other age groups with statistical difference(P <0.05).Conclusion MP and CT are easy to be detected out in summer;MP is easy to be detected out in children over 3 years old than other age groups;MP is easy to be detected out in neonates than other age groups.Fluorogenic quantitative PCR is rapid,sensitive and highly specific for the diagnosis of MP and CT infec-tions.
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Fluorogenic quantitative polymerase chain reaction(FQ-PCR) is an emerging technique with high specificity,susceptibility,automatism and has been widely used for the detection and quantification of microorganism,some genetic diseases and cancer.The design and establishment of standard FQ-PCR laboratory are required not only by gene expansion and diagnosis but also by biosafty and veracity.The aim of the paper is to introduce the structural characteristics,instruments requirement and comfort parameters of FQ-PCR laboratory.
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Objective To observe the expression level of HOXB6 gene on the differentiation and proliferation of hematopoietic stem cells(HSC) to colony forming unit-granulocyte-monocyte(CFU-GM),erythroid progenitor(CFU-E) and colony forming unite-T-lymphocyt(CFU-TL).And the proliferation procress was affected by all-transretinoic acid(ATRA).Methods 1.By the colony culture in vitro,the impact of ATRA on the CFU-GM,CFU-E,CFU-TL colony formation were surveyed;the expressions of HOXB6 gene were observed on the differentiation procress of HSC to CFU-GM,CFU-E and CFU-TL affected by ATRA on the 3rd,7th,12th.2.Real-time fluorogenic quantitative reserve transcription-polymerize chain reaction(FQ-RT-PCR) method was used to explore the possible mechanism of ATRA up-regulated to human cord blood CFU-GM,CFU-E and CFU-TL in gentic level.Results 1.HOXB6 gene had a regulatory function in the differentiation procress of hematopoiesis.During the differentiation and proliferation of HSC to colony forming CFU-GM or CFU-E,CFU-TL in vitro,the expressions of HOXB6 gene were significant positive.2.Compared with the expressions of CFU-GM and CFU-TL HOXB6 mRNA on day 3,the quantity of HOXB6 mRNA was obviously higher on day 7 and lower on day 12,respectively in each group.Compared with the expression of CFU-E HOXB6 mRNA on day 3,the quantity of HOXB6 mRNA was obviously higher on day 12 in each group.3.Compared with the HOXB6 gene of control group,the expressions of HOXB6 gene of group ATRA were up-regulated remarkable.Conclusions 1.HOXB6 gene has a regulatory function in the differentiation procress of hematopoiesis.HOXB6 gene plays an important role in the progress which can be associated with the regulating effect of HOXB6 gene on the differentiation and proliferation of CFU-GM,CFU-E and CFU-TL.2.During the differentiation and proliferation of HSC,the expression of HOXB6 gene is regular in time.3.Low concentration of ATRA(60 ?mol?L-1) can up-regulate the expression of HOXB6 gene,which confirms the theory that the normal hematopoie-tic lineage determination and maturation rely on the stable and consistent expression of HOXB6 gene.
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AIM: To establish a fluorogenic quantitative polymerase chain reaction (FQ-PCR) method for the routine examination of c-erbB-2 gene expression in breast cancer. METHODS: The c-erbB-2 standard gene was obtained by in vitro amplification of cloned c-erbB-2 fragment in plasmid PGEM-T easy vector. FQ-PCR product was detected by using a 7700 ABI PRISM sequence detector system and c-erbB-2 standard curve was obtained to quantity c-erbB-2 in unknown samples. RESULTS: “S” kinetics curve of FQ-PCR amplification was generated by relating the fluorescence signal intensity (△Rn) to the cycle number. The standard curve of c-erbB-2 was constructed by the linear relationship between the cycle threshold (ct) and the log of starting copy number. The high correlation (0.999) revealed the reliability of FQ-PCR. CONCLUSION: The FQ-PCR is a rapid, sensitive, reliable method for quantity of c-erbB-2 gene expression.
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Objective To detect the expressions of annexin 5 mRNA and Bax mRNA in the rat testis after the submandibular gland was removed. Methods On days 14,28 and 42 after the operation,the expression levels of annexin 5 and Bax mRNA in the rat testis were examined by fluorogenic quantitative RT-PCR. Results Fluorogenic quantitative RT-PCR showed that there was a 38.5% and 55.3% increase in the expression of annexin 5 mRNA and a 70.6% and 80.5% significant increase in the expression of Bax mRNA(P