Objective To study the double restriction fluorescent labeling (DRFL) method for fluorescent labeling of trace DNA samples and its effect in enhancing the pathogen detection sensitivity of microarray assays. Method SARS-CoV RNA samples were reversely transcribed and then further amplified with the restriction display (RD)-PCR and fluorescently labeled by conventional restriction labeling directly with Cy-universal primer and the novel double labeling with Cy-universal primer and CydNTP. The labeled samples were applied to the microarray with the viral probes, processed and analyzed. Results Compared with the conventional method, DRFL labeling resulted in 3. 5835 times higher fluorescent intensity of all the SARS probes on average, even though increased fluorescent intensities for different probes varied considerably. Conclusion Signal to noise ratio can be enhanced by the DRFL method which improves the sensitivity of microarray technology in trace pathogen detections.

Objective To investigate the feasibility of preparing HCV-1 diagnostic microarray probes using the technique of cDNA fragments library construction. Methods The full-length cDNAs of HCV of subtypes 1a and 1b were digested with restriction endonuclease Sau3A I, and the resulted fragments were cloned into the pMD18-T vector. Positive clones were isolated and identified by sequencing. Results A total of 57 different fragments were obtained, and sequence analysis showed that all the fragments ranging from 200 to 1000bp were specific gene fragments of HCV genotype 1, which can be efficiently used as probes in microarray prepapration. Conclusion The method of preparing microarray probes by construction of cDNA fragments library was effective, quick and simple.

Objective Based on the age of big data, the treatment mechanisms of Qishen-Yiqi(QSYQ) for myocardial infarction was focused. Methods All the data stemmed from Gene Expression Omnibus (GEO). For ensuring the efficacy genetic molecular, differentially expressed genes were discobered by Qlucore Omics Explorer (QOE). And then gene set enrichment analysis was performed by Database for Annotation, Visualization and Integrated Discovery online Gene Annotation system(DAVID) for showing the genes functions during bioprocess. In the end, the predicted genes and proteins interactions networks were demonstrated by Gene/protein interaction database (STRING). Results The main biological process involved up regulating the expression of some specil genes such as NFIL3, ARNTL, DBP, FGD4 and nuclear receptor genes like NR1D1, NR1D2 while using QSYQ treatment. In such case, BHLHE40/41 was regulated. Conclusion Traditional Chinese medicine, QSYQ, could influence the expression of genes of cytothesis, inflammatory and enzymatic activity as its effect of the molecular mechanism, in order to treat and cure the myocarditis infarction.

Objective To prepare the microarrays for joint detection of HBV and HDV. Methods The specific primers of PCR were designed with Primer Premier 5.0 program according to the conserved regions of HBV and HDV. The PCR fragments were purified and cloned into the pMD18-T vectors. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. The DNA microarray was obtained by spotting PCR products onto the surface of glass slides by robotics. Restriction display PCR (RD-PCR) was used to label the samples. Results After the sequences were aligned, we found that the products of PCR amplification were the specific gene fragments of HBV and HDV. The hybridized signals on gene chips indicated that the specificity and sensitivity of DNA microarray for joint detecting the HBV and HDV were satisfactory. Conclusion Using PCR amplification products to construct gene chips is a quick, simple and effective method for clinical diagnosis of HBV and HDV. Further application of restriction display PCR technique in labeling the sample may expedite and raise the sensitivity in multi-virus detection by microarray technology.

Objective To investigate the applieation of restriction display (RD) technique in the preparation of HCV probes of clinical genotyping microarray. Methods Restriction enzyme Sau3A Ⅰwas chosen to digest the full-length HCV cDNAs of three distinct subtypes, i.e.1a, 1b and 2a. The resultant restrictive fragments were then ligated with universal adapters. PCR primers were designed to match the universal adapters but with one "nesting" base overhanging at the 3′- end. The PCR reactions were performed by ten pairs of different primer combinations. The differential genes were separated through polyacrylamide gel electrophoresis and silver staining. The second-round PCR was performed using the isolated bands as PCR templates. The purified PCR products were then cloned into T-vectors. The recombinant plasmids were extracted from positive recombinant clones and the target gene fragments were sequenced. Results The target HCV gene fragments ranging from 200 to 900bp were isolated and sequenced, which were correlated precisely with the RFLP (Restriction Fragment Length Polymorphism) prediction. A total of 66 different fragments were obtained, averaging about 22 for each subtypes. These fragments could be further used as probes in HCV microarray preparations. Conclusion RD technique is of great value in obtaining a large number of equal sized gene probes, which provide a swift protocol in generating DNA probes for the preparation of microarrays.

Object To establish an effective plantlet regeneration system of Aconitum carmichaeli Debx. for the purpose to obtain a large number of high quality seedling in a short time. Methods Leaves in vitro were tried as the explants and cultivated in different media with the various portion of hormones. Results The medium of MS+BA 1.0 mg/L+KT 0.5 mg/L+NAA 0.2 mg/L was the most suitable one for the induction of shoots; the medium of MS+BA 1.5 mg/L+NAA 0.1 mg/L was beneficial to the propagation of shoots; the medium of MS+BA 0.5 mg/L+NAA 0.1 mg/L was good for the elongation of shoots; and the medium of 1/2 MS without any hormone was suitable for the induction of roots. Conclusion Rapid propagation of A. carmichaeli could be achieved by tissue culture and this will lead to the possibility for its seedling in the industrial production.

Graduate education is a major way for training advanced talents in the field of military preventive medicine,and the process management is the key step for training talents with high quality of theoretic creation and experimental skills.In order to cultivate excellent graduate students with distinct military especially naval characters,in recent years,we have made certain exploration and practices during the education management process,facilitating construction of educational and experimental platform,intensifying theoretic ability and lab skills,improving research project selection and design,emphasizing middle evaluation and so on.Now a new type of process management mode of graduate education in military preventive medicine with significant military character has come into being.

Objective To study the efficacy of low dose of combined angiotensin Ⅱ type 1 receptor blockade(ARB)valsartan with angiotensin-converting enzyme inhibitor(ACEI)ramipril on the expression of the gene of angiotensin Ⅱ type 1 receptor(AT1R)and type 2 receptor(AT2R)in cardiovascular vessels and brain in SHR.Methods SHRs 7-8 weeks old were received valsartan 30 mg/(kg?d),or ramipril 1 mg/(kg?d),or valsartan 15 mg/(kg?d)combined with ramipril 0.5 mg/(kg?d)by gavage for three months.SBP,LV/BW ratio,plasma angiotensin Ⅱ,plasma and myocardial NO levels were determined.The severity of myocardial interstitial fibrosis was assessed by image analysis.ACE mRNA,AT1R mRNA and AT2R mRNA expression were detected in the LV myocardium,aorta and brain by the RT-PCR.Results Combined low dose of valsartan and ramipril was shown to reduce more significantly the expression of AT1R mRNA and ACE mRNA in myocardium,aorta and brain than valsartan or ramipril monotherapy(AT1R mRNA:P

Objective: To observe the effect of metoprolol combined with trimetazidine on coronary heart disease with chronic congestive heart failure, and its effects on myocardial structure, plasma brain natriuretic peptide (BNP) and matrix metalloproteinase-9 (MMP-9). Methods: From January 2016 to November 2017, 160 coronary heart disease patients with chronic congestive heart failure in the First People's Hospital of Wenling were randomly divided into treatment group (78 cases)and control group (77 cases)through random number table method.The control group was given conventional anti-heart failure treatment.The treatment group was treated with metoprolol and trimetazidine.After 3 months of treatment, left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume(LVESV), LVEF, BNP and changes in MMP-9 were compared. Results: The total effective rate of the treatment group was 90.36%, which was significantly higher than 77.92% of the control group (χ²=5.024, P<0.05). There were no statistically significant differences in LVESV and LVEDV between the two groups before and after treatment (t=0.07, P=0.47; t=0.75, P=0.22; t=0.90, P=0.18; t=1.24, P=0.10). Compared with before treatment, the LVEF change in the control group had no statistically significant difference(P>0.05), the LVEF of the treatment group was increased[(41.38±9.26)% vs.(46.31±10.8)%] (t=-3.15, P=0.001). After treatment, the LVESV between the treatment group and control group had statistically significant difference[(124.54±16.57)mL vs.(106.36±16.44)mL](t=7.16, P<0.05). The BNP level was decreased in the treatment group after treatment[(4 036.39±696.24)ng/L vs.(3 621.78±732.57)ng/L] (t=3.73, P<0.05), while the BNP level had no statistically significant change in the control group (t=1.47, P=0.07). the MMP-9 level in the treatment group was lower than that before treatment[(396.21±97.56)ng/mL vs.(345.11±86.25)ng/mL](t=3.57, P<0.05), while the MMP-9 level in the control group had no statistically significant change (t=1.06, P=0.15). The MMP-9 and BNP levels decreased more significantly in the treatment group than those in the control group (t=3.73, 3.57, all P<0.05). Conclusion Metoprolol combined with trimetazidine in the treatment of patients with chronic heart failure can significantly alleviate clinical symptoms, reduce the expression level of MMP-9, and is safe for clinical use.Therefore, it is recommended use in clinical.

Several factors can contribute to the formation of aneurysms, including hemodynamic changes, polyarteritis nodosa, bacterial endocarditis, vasculitis, fibromuscular dysplasia, vascular malformation, and cystic medial necrosis.[1,2] Surgery is recommended for splenic artery aneurysms (SAAs) greater than 25 mm in diameter, and several surgical approaches are used, including open surgery, laparoscopic surgery, and percutaneous embolization. Laparoscopic surgery might be associated with an increased risk of pancreatic leakage compared to the open surgery approach. Open surgery without complete aneurysm resection should be preferred for patients with large SAAs in close contact with the pancreas. Here, we report a patient with two splenic artery aneurysms.