10.3969/j.issn.2095-4344.2013.24.005

Objective To establish a simple,rapid and sensitive nucleotide polymorphisms genotyping method in order to conduct the routine clinical detections under the simple laboratory condition by this method.Methods Based on the ligase-agarose gel electrophoresis,the oligonucleotide detection probes of mutational sites was designed.The detection underwent the detection probe connecting,purification and universal amplification,finally the mutation genotypes of detection sites were judged by the ap-peared bands in the agarose gel electrophoresis(AGE).With the 3 SNP sites EGFR,c.2573T>G(L858R),EGFR,c.2582T> A (L861Q)and EGFR,c.2155 G>T(G719C)in epidermal growth factor receptor(EGFR)gene as the detection objects,the plasmid template and plasma circulating DNA sample in lung cancer were performed the detection.Results The established method was easy to operate with higher specificity and sensitivity.After 20-30 cycles of PCR amplification,the genotype of detection sites was clearly estimated according to the amplification band.When detecting the mixed alleles in the heterogeneous sample,minimal 2.5%mutation alleles could be detected out.This method and the direct sequencing method could respectively detect 6 cases and 2 cases of heterozygotes mutation in the SNP site of L858R among 62 samples of lung cancer.Conclusion The established detection method for SNP genotyping is suitable to the routine mutation detection on the heterogeneous samples under the simple laboratory condi-tion.

BACKGROUND:Macrophage migration inhibitory factor (MIF) is a multi-potent cytokine that makes considerable contribution to the regulation of inflammatory response and immune response in the body. MIF rs1007888 is associated with various inflammatory diseases, but the correlation between rs1007888 and coronary heart disease in the Kazakhs of China has been rarely explored. OBJECTIVE:To investigate the relationship between rs1007888 gene polymorphisms in MIF gene and coronary heart disease in the Kazakhs from Xinjiang Uygur Autonomous Region, China. METHODS:A total of 230 Kazakh patients with coronary heart disease evidenced by coronary arteriography between December 2012 and July 2014 were recruited, and another 478 Kazak controls were free from coronary artery disease with normal angiograms. Real-time fluorescence quantitative PCR assay was used to detect the rs1007888 polymorphisms of MIF gene. Alele and genotype distributions of the rs1007888 polymorphism were compared between patients and controls. RESULTS AND CONCLUSION:Distribution of genotypes in the two groups appeared to be in Hardy-Weinberg equilibrium (P> 0.05). The alele frequencies and genotypes of MIF-rs1007888 showed no significant difference between the two groups (P > 0.05). Therefore, the genetic variation of rs1007888 in MIF gene is not associated with coronary heart disease in the Kazakhs of China.

BACKGROUND:Studies have shown that fibrin glue can promote the survival of myoblast grafts, reduce infarct size and induce neovascularization of infarct zone. OBJECTIVE:To understand the condition of revascularization of infarcted heart muscle using endothelial progenitor cells combined with degradable fibrin glue materials. METHODS:A total of 27 Sprague-Dawley rats were randomized into three groups, 9 rats in each group:non-myocardial infarction group, immediate transplantation group and 1-week post-infarction transplantation group. Then, these three groups were sub-grouped into two groups, respectively:endothelial progenitor cells+fibrin glue group (experimental group) and fibrin glue group (control group). At 3 and 8 weeks after transplantation, the rats were sacrificed in each group. The revascularization and function of infracted heart muscle were observed by microscope, immunohistochemistry and echocardiography. RESULTS AND CONCLUSION:Under the microscope, there were some lax connective tissues between the heart and chest in the experimental groups, but no difference existed between the experimental and control groups. The heart structure was normal relatively and difficult to be distinguished between the experimental and control groups histological y and immunological y, and there was no angeioma, vascular malformation and tumor. The number of revascularization of heart muscle showed no difference between experimental and control groups as wel as between different experimental groups. Additional y, there was no significant difference in cardiac function between experimental and control groups. Although there are no positive results of endothelial progenitor cells, we wil modify and improve the strategy and believe that the celldelivery system is of benefit and efficacy.

Objective To evaluate the stability of U6 and let-7a as internal reference genes of miRNAs in RTqPCR by using femoral head samples of cartilage tissue from inbred DA rats.Methods Total RNA was extracted from femoral head cartilage tissues of female DA rats at three different time points,i.e.at birth (D0),ablactation (D21) and maturation (D42).The expressions of different miRNAs (miR-1,-25,-26a,-140,-146a,-150,-181a,-195,-223 and-337) were detected by RT-qPCR using U6 or let-7a as the internal reference.The two sets of miR expression were compared with the results from Solexa sequencing in our pioneer work to evaluate the stability of the two internal references.Results The relative values of U6 (P =0.045) and let-7a (P =0.021 5) revealed significant difference in the D42 sample.Both in U6 and let-7a systems,miR-26a,-140,-223,and-337 showed a similar tendency in expression and quantification but miR-1 and-146a did not have significant differences.miR-25,-150,-181a and-195 differed significantly (P＜0.05).Comparison of absolute quantification results between the two generations' sequencing showed that let-7a is more stable than U6.Conclusion Let-7a is more suitable to be used as the internal reference gene in RT-qPCR for miRNAs in cartilage tissue.

BACKGROUND:Cyclin A2 is a key regulator of cellcycle. Location and expression of cyclin A2 in neonatal mouse myocardium is not clear.
OBJECTIVE:To observe the location and expression of cyclin A2 in neonatal mouse cardiomyocytes and its relationship with the exit of cardiomyocytes from cellcycle.
METHODS:Neonatal mice were kil ed to take myocardial tissues at 0, 3, 7, 14 and 28 days after birth. Western blot were used to detect the expression of cyclin A2, proliferating cellnucleus antigen and Phospho-histone H3. Immunohistochemitry detection was used to detect the location of cyclin A2 and expression of proliferation cellnucleus antigen at different time after birth.
RESULTS AND CONCLUSION:Western blot showed the decrease of cyclin A2 after birth til disappeared at day 4 (P=0.001). Cyclin A2 located mainly in the nucleus after birth and exported to the cytoplasm at day 14, and basical y disappeared at day 28. Proliferating cellnucleus antigen showed gradual y decreased tendency after birth. Mitosis specific marker, Phospho-histone H3, exhibited a gradual decrease after birth, which was consistent with cyclin A2 in expression intensity.

Provisional restorations are beneficial to final restorations. However, the importance of provisional restorations is often overlooked because they are designed to function for a limited period. This article summarizes the clinical values and two common materials of provisional restorations.

Objective:To explore the relationship between psychological resilience,professional identity and employment pressure of nursing students with different educational levels,and analyze its influencing factors.Methods:A total of 208 nursing students with different educational levels were investigated by using the general information questionnaire,the employment pressure questionnaire of nursing students,the psychological resilience scale and the professional identity questionnaire.The correlation among psychological resilience,professional identity and employment stress was analyzed by Pearson correlation analysis.Results:The total score of professional identity,psychological resilience,and employment stress was(54.46±12.34),(57.62±18.66),and(98.30±20.25),respectively.There was no significant difference in professional identity score and psychological resilience score among nursing students with different educational levels(P＞0.05).There was no significant difference in the employment stress scores between nursing students upgrading from a junior college and undergraduate and junior college nursing students(P＞0.05),while the employment stress score of junior college nursing students were significantly higher than that of undergraduate nursing students(P＜0.05).Pearson correlation analysis showed that the total score of employment stress was positively correlated with the total score of psychological resilience(r=0.252),and the total score of professional identity(r=0.312)(P＜0.01).Conclusions:The psychological resilience of nursing students is at the average level,professional identity is low,and the employment pressure of junior college nursing students is higher than that of undergraduates.Targeted interventions should be developed to improve the psychological resilience and professional identity of nursing students,and reduce the employment pressure level of junior college nursing students.

Objective To analyze the research trends and hotspots of tenecteplase and provide a reference for clinical research and rational drug use in China.Methods A comprehensive search was conducted on tenecteplase studies in the Web of Science database from its inception to April 2024.CiteSpace 6.1.R6 software was used to analysis authors,countries/regions,research institutions,keywords,and co-cited references.Results A total of 673 articles were included in this study.The annual publication volume showed an overall upward trend,rapidly increasing in the past three years.The United States,Canada,and Australia were the top 3 countries in terms of publication volume in this field.Among the top 10 institutions in publication volume,9 were from above 3 countries.The research hotspots were the efficacy and safety of tenecteplase in patients with acute myocardial infarction and acute ischemic stroke.In recent years,research hotspots have mainly focused on the field of acute ischemic stroke.Conclusion The application of tenecteplase in the field of acute ischemic stroke is currently a research hotspot.However,there is still a gap between the research level in China and that in developed countries such as the United States.Future research hotspots may still focus on the application of tenecteplase in patients with different types of acute ischemic stroke,suggesting that scholars should pay close attention to this area.

Objective:To construct the standardized preventive nursing training scheme for venous thromboembolism (VTE) , so as to provide a basis for hospitals to carry out the standardized preventive nursing training scheme for VTE.Methods:Under the guidance of the National Medical Quality and Safety Improvement Goal, the standardized preventive nursing training scheme for VTE was initially developed through preliminary investigation, literature review and group discussion. Using the convenient sampling method, a total of 20 experts were selected from March to June 2022 for two rounds of Delphi expert consultation. After modification, the VTE standardized preventive nursing training scheme was finally determined.Results:In the two rounds of correspondence, the effective response rates of the questionnaire were 90.00% and 100.00%, respectively. The expert authority coefficients of two rounds were both 0.95 and Kendall coordination coefficients were 0.20 and 0.14, respectively ( P<0.01) . After two rounds of expert consultation, a standardized preventive nursing training scheme for VTE was finally established, which included 4 primary indicators, 12 secondary indicators and 78 tertiary indicators. Conclusions:The VTE standardized preventive nursing training scheme constructed in this study has strong reliability and scientific nature, which can provide a basis for hospitals to carry out VTE standardized preventive nursing training scheme.