Suppression subtractive hybridization(SSH),which is feasible and highly specific,is one of the most efficient methods for cloning differentially expressed genes.It has been successfully applied in the study of tumors,stem cells,etc.

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 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 evaluate the effect of direct percutaneous transluminal coronary angioplasty (PTCA) in early stage in acute myocardial infarction (AMI) by acoustic densitometry (AD) technique Methods 40 patients with AMI were divided into 2 groups: 18 patients with direct PTCA (group A),22 patients with medical treatment (group B) The results of 2 D echocardiography and AD examination were studied on both groups at 7 days after admission Results There was no statistical difference in routine echocardiography between the two groups The results of AD examination showed: cyclic variation of integrated backscatter (CVIB) in group A was (4 2?1 3)dB, it was significantly higher than that in group B (2 5?2 4)dB ( P 0 05) Conclusion AD technology can reflect the changes of tissue characteristics in reperfused myocardia early It is a new technology for evaluating the effect of interventional therapy, predicting the improvement of cardiac function, and assessing the myocardia viability

Objective To explore the effects of osteoblasts on the recovery of hematopoiesis and angiogenesis in acute irradiation injury mice.Methods The femurs of 18 male BALB/c mice were used to prepare the bone marrow osteoblasts, and the rest mice were divided into 3 groups as normal group, saline group and osteoblast group.The mice in normal group received no treatment, and the other two groups were received 6.0 Gy 60Co γ-ray irradiation.After irradiation each mouse of osteoblast group was administered with 2 × 106 osteoblasts through tail vein injection, and equal volume saline was given to each mouse of saline group by the same way.The following factors were measured at 7, 14, 21 d after irradiation, they were the counts of peripheral blood cells and bone marrow mononuclear cells ( BMMNC ) , the percentage of CD34 + cells in BMMNC, the histology changes and micro vascular density (MVD) of bone marrow tissue.Results The counts of peripheral blood cells, BMMNC and hematopoietic tissue area in osteoblast group were higher than those in saline group.The percentage of CD34 + cells in BMMNC and the MVD of bone marrow in osteoblast group were also higher than those in saline group at 7, 14, 21 d after irradiation ( t = 2.46 - 64.51, P ＜ 0.05 ).Conclusions Osteoblasts could significantly promote the recovery of hematopoiesis and angiogenesis in mice after acute irradiation injury.

Objective To explore the effects of cotransplantation with osteoblasts on hematopoietic reconstitution in mice after bone marrow transplantation (BMT). Methods The typical model of syngeneic BMT was established. 18 Balb/c mice were used to prepare the bone marrow nuclear cells and osteoblasts for BMT. The 42 Balb/c mice were randomly divided into 3 group:normal group (6 mice, without any treatment), the single BMT group ( 18 mice, given 2 × 106 bone marrow nuclear cells/each mouse) and the cotransplantation group of HSC with osteoblaats (18 mice,given 2 × 106 bone marrow nuclear cells and osteoblasts/each mouse). The following factors were measured on day 7, 14, 21 after BMT: peripheral blood cells, bone marrow mononuclear cells (BMMNC), the percentage of CD34+ cells in BMMNC (assayed by flow cytometry), the hematopoietic tissue changes (detected by HPIAS-1000 image analysis system) and micro vascular density (MVD) of bone marrow tissue (with immunohistochemistry). Results The levels of periphral WBC, RBC, PLT, BMMNC in the contransplantation group were higher than those in the single BMT group (P＜0. 01 or P＜0. 05). In the contransplantation group, the percentage of CD34+ cells in BMMNC, the hematopoietic tissue area and the MVD of bone marrow were also higher than the single BMT group on the 7th, 14th, 21st day after BMT(P＜0.01 or P＜0.05). Conclusion Cotransplantation with osteoblasts could significantly promote hematopoietic reconstruction in mice after BMT. Cotransplantation may represent a promising means of achieving higher engraftment rate after BMT.

Objective To develop a better probe preparation method of gene chips for the further gene chips fabrication and its application in the gene expression. Method Using human leukemia K562 cells as material,applying a new kind of technology for gene differential display restriction display(RD PCR),we isolated a lot of cDNA fragments as probes of gene chips. Results This method has overcome the shortcoming of more pseudopositives in DD PCR,and was easy to be manipulated.Compared with the chip fabricated with full cDNA probes,the probe length is comparatively homogeneous,and the hybridization dynamics is apt to be controlled.Conclusion\ RD PCR technology could provide a new strategy for the probe preparation of gene chips.\;[

Objective:To research the TLC identification method for Longdan Xiegan pills( honey pills) . Methods:TLC was used in the identification. The samples were extracted by 70% methanol with a heating reflux method, and then extracted by the agents with dif-ferent polarity, including petroleum ether, ethyl acetate and butanol. The petroleum ether part was detected by fluorescence at 365nm for Angelica sinensis, and 1% vanillin-sulfuric acid color reaction was used to detect Alisma orientale. The ethyl acetate part was determined by fluorescence at 365nm for Scutellaria baicalensis, and the butanol part was detected with chloroform-methanol-water (30∶12∶3) as the developing solvent for Bupleurum and Glycyrrhiza, and with acetone-ethyl acetate-water (6∶6∶1) as the developing solvent for gentiopi-croside, geniposide and liquiritin. Results:The developed TLC spots were clear with good separation, high specificity and promising re-producibility. Conclusion:The method can be exactly used in the qualitative identification and quality control of Longdan Xiegan pills ( honey pills) .

Objective To analyze the complications,treatments and prognosis of simultaneous pancreas-kidney transplantation,especially on surgical complications and treatments.Method The causes and outcomes of surgical treatment in 70 cases of simultaneous pancreas-kidney transplantation performed between Dec.1999 and June 2012 were retrospectively analyzed in our center.Result Sixteen patients (22.9％) underwent one or more reoperations.The causes for reoperation were as follows:2 cases of hematuria,4 cases of abdominal hemorrhage,4 cases of abdominal infection,4cases of pancreatic thrombosis,2 cases of renal graft's artery rupture,1 case of renal allograft rupture,1 case of intestinal fistula,and 1 case of pancreatic fistula.Eight pancreas grafts were lost in the first year.Pancreatectomy was performed on the other 5 cases:4 cases of pancreatic thrombosis,1 case of intestinal fistula,accounting for 43.8％ of the patients subject to reoperation.The recipients,kidney,pancreas survival rate in reoperation group at 1 year was 87.5％,75％,and 56.3％ respectively; and that in control group at 1 year was 98.1％,98.1 ％,and 98.1 ％ respectively.There was significant difference in kidney survival rate (P＜0.01,chi-square =6.79),and pancreas survival rate (P＜0.01,chi-square =17.47) between two groups.Conclusion Although simultaneous pancreas-kidney transplantation provides a successful and effective treatment for diabetics with end-stage renal disease,surgical treatment due to complications is still an important factor in short-term survival on the grafts.

Objective To evaluate right ventricular(RV)function by tissue Doppler imaging(TDI)and Tei index in patmnts with coronary artery disease(CAD).Methods One hundred twenty patients who underwent coronary angiography(CAG)were enrolled.Based on clinic evidences,such as CAG,disease history,symptom,ECG,and myocardial enzymes,all patients were divided into three groups:control group,angina group and myocardial infarction(MI)group.TDl was performed on the lateral and septal tricuspid annular.RV Tei index was obtained either from the pulse-wave Doppler(PW-Tel)and from TDI(TDI-Tei).Results There were significant differences in RV PW-Tei,TDI-Tei,Ea,and Ea/Aa among three groups.PW-Tei was 0.363±0.086.0.463±0.078 and 0.569±0.066,and TDI-Tei was 0.400±0.058,0.520±0.078 and 0.628±0.059 for the control group,angina group and MI group,respectively.Ea and Ea/Aa were gradually decreased following the severity of CAD.Sa was no difference between the control group and angina group,while Sa was much lower in mvocardial infarction group than in other groups.RV PW-Tei had a good correlation with the lateral and septal TDI-Tei.The lateral and septal TDI-Tel were significant correlated with motion velocities.Conclusions TDI can be used to comprehensively evaluate RV function in patients with CAD.TDI-Tei can sensitive reflect the damage level of overall RV function in patients with different degree CAD.