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 study the expression of interleukin-18(IL-18)during the progression of renal interstitial fibrosis in rat kidney after unilateral ureteral obstruction(UUO)and the effect of Shenfu injection.Methods The obstructive nephropathy model was established by unilateral ureteral obstruction(UUO).Fifty。Six rats were randomly assigned into shame operation group,operation group(UUOgroup)and treatment group(UUO+Shenfu).After 7 and 14 days,the renal function and histopathological changes were evaluated.Immunohistochemistry was used to examine the expression of IL-18 in renal tissue.Results In comparison with the shame opeartion group,the operation group showed obvious renal interstitial fibrosis.And the expression of IL-18 increased signifieantly(P<0.05).Compared with the operation group,the degree of interstitial fibrosis was obviously ameliorated in the treatment group,and the expression of IL-18 decreased significandy after treatment for 7 days(P<0.05),and decreased more after treatment for 14 days(P<0.05).Conclusions Shenfu injection may protect renal function by decreasing the expression of IL-18 in the progression of renal interstitial fibrosis.

Objective To explore the technology of excision,and finishing on donative pancreas during combined pancreas-kidney transplantation.Methods We successfully harvested multiple abdominal organs together on 40 cases.Wide surgical exposure was obtained.Cannulas were placed for in situ cooling in portal vein and abdominal aorta,and flushed with HC-A (2000 ml) and UW (1000 ml) with the pressure being 10 cm H2O.The intestine was flushed with 0～4 ( normal saline (1000 ml) and metronidazole (200 ml),the liver,kidney,pancreas,spleen and duodenum were en bloc excised and isolated,and the pancreas and kidney were pruned.Results Excision of donative abdominal organ was successfully performed on all 40 cases.The en bloc warm ischemic time was 3.2 min (2～5 min).The cold ischemic time of pancreas was 10.6 h (8～15 h).The cold ischemic time of kidney was 8.5 h (4～16 h).Post-operation mean withdrawal-insulin time was 9.5 days (4～17 d),FFG 6.7 μmol/L (4.4～10.7 μmol/L),GHbA1c 4.4 ％ (4.1 ％～4.7 ％).Creatine was 87.2 (56～121) μmol/L one month after operation.There were 2 cases of DGF after operation,and the creatine level was returned to the normal within one month after operation.Conclusion Technology of excision,preservation and finishing on donative pancreas for combined pancreas-kidney transplantation was one of the key points for successful transplantation.

Objective To study the effect of atorvastatin on serum cystatin C and urinary microprotein in patients with early diabetic nephropathy (DN).Methods Sixty-eight cases of early DN were divided into control group and observation group with 34 cases each by random digits table,the control group was treated with the conventional treatment,the observation group was treated with atorvastatin on the basis of conventional treatment,then blood lipids,serum cystatin C,urinary albumin excretion rate (UAER),microalbuminuria( MAU ),α1-microglobulin(MG),β 2-MG between the two groups were compared.Results After intervention,the levels of total cholesterol (TC),triacylglycerol (TG),serum cystatin C,UAER,MAU,αt 1-MG,β 2-MG were significantly decreased compared with those before treatment in the observation group [ (4.32 ± 1.26) mmol/L vs.(5.65 ± 1.38 ) mmol/L,( 1.67 ± 0.64) mmol/L vs.(2.53 ± 0.96 ) mmol/L,( 1.29 ± 0.38 ) mg/L vs.( 1.74 ± 0.51 ) mg/L,(61.09 ± 18.45 ) μ g/min vs.( 86.42 ± 21.34) μ g/min,( 5.73 ±4.81) mg/L vs.(23.16 ±9.73) mg/L,(1.41 ± 1.21) mg/L vs.(4.76 ± 1.24) mg/L,(1.21 ±0.13) mg/L vs.(2.58 ± 0.26) mg/L ] (P ＜ 0.01 or ＜ 0.05 ).In the control group,the levels of TC,TG,serum cystatin C were lower than those before treatment,but there were no significant differences (P＞0.05),but the levels of UAER,MAU,α 1-MG,β 2-MG had significant differences compared with those before treatment (P ＜ 0.01or ＜ 0.05 ) and the same time of the observation group (P ＜ 0.05 or ＜ 0.01 ).Conclusion Atorvastatin can significantly reduce serum cystatin C and urinary micro-protein levels in patients with early DN,which plays an important role in kidney protection.

Objective To analyze the complications, treatments and prognosis of simultaneous pancreas-kidney transplantation. Methods Forty cases of simultaneous pancreas-kidney transplantation performed between Dec. 1999 and Jan. 2010 in our center were retrospectively analyzed. Results Regarding surgical complications, 4 cases had severe hematuria after operation,which needed clinical intervention, including 1 patient receiving catheterization in duodenum to stop bleeding. Two patients were treated with continuous bladder irrigation, and the remaining one received surgical haemostasis because of donor's duodenum and bladder anastomotic artery hemorrhage.Abdominal hemorrhage occurred in 4 patients, including pancreatic hemorrhage in 3 cases and duodenal muscularis hemorrhage in one case. All of them received surgical treatment for hemostasis.Abdominal infection occurred in 8 cases: one died of multiple organ failure, 2 cases were cured after drainage of abscess, 2 cases underwent surgical removal of abscess, and 3 cases were cured after antibiotic therapy. In one case of postoperative anastomotic leakage, pancreas was resected. Four cases of postoperative ileus were cured by continuous clysis with traditional Chinese medicine. Seven cases had pulmonary bacterial infections, including one cases associated with fungal infection. They were cured by the anti-infective treatment. Other complication included poor healing in 5 cases and urinary infection in 2 cases. After combined simultaneous pancreas-kidney transplantation, 10 patients received reoperation because of surgical complications (14 operations). The re-operation rate was 25 %, including 2 patients (4 operations) for hematuria, 4 patients for abdominal hemorrhage, 2 patients (3 operations) for abdominal infection, 1 patient for pancreatic venous thrombosis, 1 patient for anastomotic leakage, and 1 patient for pancreatic fistula. Conclusion Although simultaneous pancreas-kidney transplantation provides a successful and effective treatment for diabetics with endstage renal disease, surgical complication is still affecting the pancreas and kidney grafts after transplantation.