Respiratory virus remains to be an important pathogen of respiratory disease in children.The disease can occur in all age groups, especially in young children.Most viral infections have a good prognosis, but special viruses still cause great harm to the health of children.Respiratory viral therapy includes symptomatic therapy, broad-spectrum antiviral drugs, drugs that directly target the viral replication cycle, drugs that attenuate the inflammatory response, and anti-viral nanodrugs.New antiviral drugs are urgently needed to develop.The repurposing of the existing therapeutic agents previously designed for other virus infections is also an effective way.The treatment of respiratory virus infection has become an important topic in clinical research.

Objective This study was designed to study the effect of Diltiazem on patency rate of arteriovenous anastomosis in rat and how it works.Methods 24 SD rats were divided into control group and experimental group,12 rats in each group.Experimental group rats were gavaged with Diltiazem after vascular anastomosis.Control group rats were gavaged with water.By comparing the patency rate and the thickness of artery to make sure whether Diltiazem will affect the patency rate.;By comparing the clotting time,prothrombin time,artial thromboplastin time,and serum thromboxane B2 levels to explore the pathway of diltiazem.Results The patency rate was 75％ in the experimental group and 25％ in control group.Compared with the control group,experimental group venous blood vessels in the film segment was significantly thicker,clotting time was prolonged,TXB2 levels in blood was decreased,the differences were statistically significant(P ＜ 0.05).There were no significant difference in prothrombin time and partial thromboplastin between two groups (P ＞ 0.05).Conclusion Diltiazem can inhibit the secretion of TXB2,antagonize the effct of antiplatelet,and increase the patency rate of vascular anastomosis in rats.

Totally 11 subjects with diabetic foot have been treated by umbilical cord blood mesenchymal stem cell (UCB-MSC) transplantation from November 2006 to October 2008 at the Chengyang People’s Hospital of Qingdao City. The course of disease was 6-18 years. Following over 6-months drug treatment and/or scaffold implantation, vascular bypass outcomes were bad. Of these patients, there were foot and/or lower limb cooling, pain, intermittent claudication and rest pain in 7 cases, abnormal color in foot skin in 7 cases, foot ulcer in 6 cases, gangrene in 4 cases, and concurrent infection in 2 cases. UCB-MSCs were obtained from healthy pregnant women, and supplied by Beike, Shenzhen, China. Following epidural or lumbar anesthesia, UCB-MSC suspension [cell number (1-6)?1011/L, 0.3-0.5 mL per point] was injected into affected lower extremity through multipoint muscles, with a distance among each point was about 3 cm ? 3 cm. Demixing injection could be performed in regions with plenty of muscles. At 1-4 weeks following transplantation, foot pain in 9 cases got amelioration or easement and cold sensation in 10 cases improved markedly. At 4-16 weeks after transplantation, foot ulcers were better and ulceration area reduced in 4 cases, gangrene area in 2 patients reduced and avoided amputation. 1 limb was amputated but with lowered level of amputation. After 12-24 weeks, 6 patients with intermittent claudication improved obviously, ankle-brachial index (ABI) increased in 3 cases, angiography showed an increase in visible collateral vessels in 3 patients. Transplantation of UCB-MSCs might be a simple, safe and effective method for patients with lower limb ischemia. It can effectively increase blood flow of lower limbs, promote ulcer healing and decrease amputation rate and amputation level.

To study the pharmacokinetics of ginsenosides Rg1 and its metabolites after iv and oral administration in Wistar rats, the LC-MS/MS method was selected to determine ginsenosides Rg1 and its metabolites in plasma and their pharmacokinetic parameters were calculated. After oral administration of ginsenosides Rg1 to rats, ginsenosides Rg1, Rh1, F1 and protopanaxatriol (Ppt) could be detected in plasma. Their Tmax were 0.92, 3.64, 5.17, and 7.30 h, respectively; MRT were 2.68, 5.06, 6.65, and 5.33 h, respectively; AUC(o-t), were 2 363.5, 4 185.5, 3 774.3, and 396.2 ng x mL(-1) x h, respectively. After iv administration of ginsenosides Rg1 to rats, ginsenosides Rg1, Rh1 and FI could be detected in plasma. Their T1/2betaS were 3.12, 5.87, and 6.87 h, respectively; MRTs were 1.92, 5.99, and 7.13 h, respectively; AUCo-tS were 1 454.7, 597.5, and 805.6 ng x mL(-1) x h, respectively. So, it can be concluded that after oral administration, the amounts of metabolites were higher than the prototype in vivo, and the distribution and elimination of the metabolites were relatively slow. After iv administration, the amount of prototype were higher than that of the metabolites in vivo, and the distribution and elimination of the metabolites were relatively slow.

BACKGROUND:The creep characteristics of cancellous bone from necrotic femoral head are important for clinical artificial joint replacement.Therefore,it is necessary to study mechanical properties of cancellous bone at 45°direction.OBJECTIVE:To compare creep properties of normal femoral head and necrotic femoral head at 45°direction based on three-parameter model established creep equation METHODS:A total Of 8 normal and 8 necrotic femoral heads were used.The cancellous bone was harvested at 45° and subjected to creep test on electronic universal testing machine.With simulated temperature field of human body temperature at36.5℃.stress was imposed on the samples by an increase of 5%/s for 7 200 seconds.100 experimental data were collected,and stress relaxation equation was calculated using three-parameter model.RESULTS AND CONCLUSION:The creep curve of the femoral head of normal and necrotic changes was exponential relation.Changes were fast in the first 600 seconds,and strain increased slowly with time,finally entered into balance stage.7 200 s creep of cancellous,bone from necrotic femoral head was less than normal femoral head cancellous bone.Three-parameter model calculation is simple and can well fit the creep of changes in the femoral head.The establishment of such idealized equation quantitatively shows poor visco-elasticity of the necrotic femoral head.

Objective To study a functional variable fragment of heavy chain(VH)antibody against the terminal protein(TP)region of hepatitis B virus(HBV)polymerase introduced by human immunodeficiency virus Tat protein transduction domain(TAT)and the inhibitive activity of TAT-VH on the replication of HBV in vitro.Methods The gene encoding TAT-VH was cloned into prokaryotic expression vector pET28a(+).Recombinant plasmid was transduced into E coli BL21(DE3)LysS,then the protein was expressed and purified.The purified TAT-VH fusion protein was added into HepG2.2.15 cell culture.The transduction efficiency was evaluated by indirect fluorescence assay(IFA).The cytotoxicity of TAT-VH was detected by Methabenzthiazuron(MTT)assay.HBV DNA level in HepG2.2.15 cell culture was measured using quantitative polymerase chain reaction(PCR).The data were analyzed by one-factor analysis of variance and t test.Results TAT-VH fusion protein was successfully expressed and purified.It was confirmed by IFA and MTT assay that TAT-VH was introduced into HepG2.2.15 cells and the cell growth was not affected.The level of HBV DNA in supernatant of HeDG2.2.15 cell culture with 5 000 nmol/L TAT-VH was(1.211±0.132)lg copy/mL,which was significantly lower than control group[(5.325±0.041)lg copy/mL,t=72.91,P＜0.05].Meanwhile,the level of intracellular HBV DNA was(3.521±0.411)lg copy/mL,which was significantly lower than control group[(8.532±0.132)lg copy/mL.t=28.41,P＜0.05].Conclusion The HBV replication is inhibited by anti-TP TAT-VH antibodies in vitro,which provides valuable experimemal basis for developing therapy of HBV infection with intracellular antibody.

Objective To detect 14 genes including fibrillin-1(FBN1) and so on mutations in 17 patients with Marfan syndrome(MFS) and family members of 2 patients and to investigate the correlation between FBN1 gene mutation and MFS.Methods Genomic DNAs were extracted from whole blood sample of 17 patients and 43 family members.After DNA samples were amplified by polymerase chain raction(PCR), we used capture panels to get target genes which would be sequenced by Illumina HiSeq2500 Analyzers(Illumina, SanDiego, USA).The target genes included ACTA2、CBS、FBN1、FBN2、MYH11、COL3A1、SMAD3、TGFBR1、TGFBR2、MYLK、MSTN、COLA2、TGFB2 and SLC2A10.The results of sequencing would be compared with multiple databases, including NCBI dbSNP, HapMap, 1000 human genome dataset and database of 100 Chinese healthy adults, to find gene mutation.Finally, these mutations would be validated using conventional Sanger sequencing methods.Results A total of 10 FBN1 mutations and 1 actin alpha2(ACTA2) mutation in 17 patients were identified, of which 8 FBN1 mutations and 1 ACTA2 mutation were novel.One FBN1 mutation was underwent family investigation and we found in this family, all patients had this mutation and others did not have it.Conclusion Missense mutation of c.7280G>A in the 59th exon of FBN1 gene is new pathogenic mutation for MFS.The other 8 novel mutations may be the pathogenic factors of MFS.

Objective To study the relationship between expression of cyclooxygenase-2 (COX-2) and vas-culax endothelial growth factor (VEGF) with liver sinusnidal capillarization (LSC) of chronic hepatitis B (CHB). Methods We studied liver biopsies from 200 patients with CHB COX-2 ,VEGF immunohistochemical stain were ob-served to accomplish relationship between expression of COX-2,VEGF and LSC. Results LSC occupy above 80% in the group. There were manifestation in mild-LSC (focal) , middle-LSC (sheet-shape) and severe-LSC (widespread). Electron microscope shown the laceration in the endothelium of sinuses and formation of basal lamina and budding for-mation lumen of blood vessel and fat-storing cell convert myofibroblast. Expression of COX-2, VEGF, Co-Ⅳ and retic-ulum, collagen and elastic fibers with mild or severe in LSC is manifest locking relate. Conclusion Increased ex-pression of COX-2 ,VEGF in liver tissue of CHB may facilitate LSC and hepatic fibrosis.

Objective To explore the related factors and diagnostic criteria of perioperative myocardial infarction (PMI) after on-pump coronary artery bypass grafting(CABG).Methods 258 CABG patients were selected.The cardiac troponin-I in immediately,6,12,24,and 48 hours after surgery were measured.95 percentile (P95) was used as the boundary,any measured value greater than P95 was identified PMI,as group I(13 cases),the rest as group II(245 cases).The age,sex,cardiopulmonary bypass time,aortic corss-clamp time,LV ejection fraction,left ventricular end diastolic diameter,grafted vessels,left anterior descending coronary artery occlusion,recent myocardial infarction (<3 months),severe complex coronary artery disease and other risk factors that may lead to PMI were analyzed.The data were analyzed using Student's test for continuous variables and the χ2 test for discontinuous variables.Results P95 value was 3.47,the cardiopulmonary bypass time(t=3.268,P<0.05),aortic corss-clamp time(t=2.047,P<0.05),severe complex coronary artery disease between the two groups had statistically significant difference (χ2=19.846,P<0.05).Conclusion cTnI>P95 (3.47) indicates that the myocardium injury is serious,cardiopulmonary bypass time,aortic corss-clamp time and severe complex coronary artery disease are associated with PMI in patients undergoing CABG.

AIM　To study the pharmacokinetics and bioavailability of clinafloxacin in rats. METHODS　The drug concentration was determined by HPLC. The main pharmacokinetic parameters were obtained by 3P87 program. An RP-C18 was used as the stationary phase. The mobile phase was a mixture of acetonitrile-0.05 mol*L-1 citric acid triethylamine (pH 2.5) (20∶80). The flow rate was 1.0 mL*min-1. The UV absorbance detector was set at 300 nm. RESULTS　A good linearity was obtained from 0.03-20 μg*mL-1 of clinafloxacin in rat plasma with γ=0.9998. The plasma concentration-time curve of clinafloxacin conformed to one compartment open model. After ig administration of 50 mg*kg-1 and 100 mg*kg-1 dose of clinafloxacin in six rats, mean Cmax and AUC values increased in proportion to dose. Mean T1/2 appeared to be independent of dose. Mean AUC was 65±6 and 27±4 μg*h*mL-1 respectively after iv and ig adminostration of 100 mg*kg-1 dose. The extent of bioavailability (F) of clinafloxacin was 42%. CONCLUSION　The results of the pharmacokinetic study of clinafloxacin showed that it exhibited first order kinetic characteristics and the bioavailability is low.