Objective To establish a gene diagnosis assay for spinal muscular atrophy(SMA) in children. Method Analysis of the survival motor neuron (SMN) gene in 19 SMA patients and in 21 normal controls were performed by using polymerase chain reaction - fragment length polymorphism (PCR - RFLP) method. Result Deletion of exon 7and 8 in SMNt gene were found in all 19 SMA patients, while no such changes were found in normal controls. Conclusion The SMNt gene exon 7 and 8 examine can be applied to SMA gene diagnosis, and the PCR- RFLP method have higher sensitivity and particularity to the SMA diagnosis.

Objective To evaluate intra-and interatrial synchronization in patients with paroxysmal atrial fibrillation (PAF) by dual gate Doppler and discuss the predictive value of atrial asynchrony for detecting PAF.Methods Twenty-nine patients with PAF as AF group and 24 healthy volunteers as control group underwent echocardiography.The time interval of the onset of a'wave between mitral annular septal and lateral site was T1 by dual gate Doppler,which was used to evaluate left atrial synchronization.The time intervals from the onset of a'wave at tricuspid annular right ventricular free wall site to a'wave at tricuspid annular septal site and mitral annular lateral site were T2 and T3,which were respectively used to evaluate right atrial and interatrial synchronization.The greater time interval predicted the worse synchronization.Compared these indexes between two groups,the most valuable index was obtained and combined with left atrial volume index (LAVI) to predict PAF,its optimal values were found out by ROC curve.Results ①There were no significant differences in many other basic indexes between two groups (P ＞0.05).However,compared with control group,the ratio of early diastolic transmitral flow velocity (E) and mitral annular velocity of septal site (e'),the diameters and volume indexes of atria were significantly increased in AF group.②T1,T2 and T3 (standardized before and after) increased significantly in AF group (P ＜0.01).Standardized T1 had the largest area under curve (AUC) for detecting PAF.The AUC of standardized T1 and LAVI were 76.6％ and 84.1％.Standardized T1 ≥29.55 ms and LAVI≥ 21.60 ml/m2 were combined for detecting PAF (sensitivity 82.8 ％,specificity 79.2％,AUC 87.1％).③ Standardized T1 was positively correlated with age,left atrial systolic diameter,LAVI,RAVI and E/e'(P＜ 0.05).Conclusions Patients with PAF have worse intra-and interatrial synchronization and left atrial asynchrony can provide predictive value for detecting PAF.Dual gate Doppler is expected to be a new technique for evaluating intra-and interatrial synchronization accurately and detecting the risk of AF.

Objective: We observed the influence of recombinant human growth hormone(rhGH) on protein metabolism in hepatocirrhotic rats and investigated mechanisms. Methods: Ninety rats were randomized into three groups(n=30 for each group): group A(normal rats), B(cirrhotic rats) and C(cirrhotic rats receiving rhGH). Rats in group C were subcutaneously given rhGH with a dosage of 1 U/(kg?d) for up to 1 week. The plasma concentrations of albumin(ALB), prealbumin(PA), transferrin(TF) and insulin-like growth factor-1(IGF-1) were detected on experimental day 14. And liver biopsy was taken to examine the expression level of albumin mRNA with TR-PCR. Results: The albumin, prealbumin, transferrin and IGF-1 levels in groups A and C were significantly higher than those in group B. The expression level of albumin mRNA in grup C was significantly higher than that in group B. Conclusions: The rhGH can improve the level of IGF-1 in plasma and improve the expression of albumin mRNA. Moreover, it may significantly improve the synthesis of protein and attenuate hypoproteinemia in cirrhotic rats.

OBJECTIVETo established the rapid tissue propagation system of Epimedium wushanense, in order to provide theoretical basis for industrialized seed cultivation.

METHODTiller buds E. wushanense were used as explants, with MS, B5, WPM as basic media, and added with different concentrations of plant growth regulators such as 6-BA, NAA and GA3, in order to conduct a systematic study on induction and propagation conditions for tiller buds.

RESULTThe suitable method for sterilizing bud was to disinfect with 75% ethanol for 30 s, and then treated with 0.1% HgCl2 for (4 + 2) min for consecutively twice, which could control the pollution rate below 5% and the survival rate above 75%. The optimal medium for bud induction was WPM + 6-BA 2.0 mg x L(-1) + NAA 0.1 mg x L(-1) + GA3 0.5 mg x L(-1), with the induction rate of 75.5%; meanwhile, the basic medium and 6-BA showed significant effect on the induction rate. The propagation medium suitable for buds was MS +6-BA 2.0 mg x L(-1) + NAA 0.5 mg x L(-1), with the propagation rate of 3.3. The optimal growth of rooting medium was 1/2 WPM + IBA 0.5 mg x L(-1) + activated carbon which (0.05%), with the rooting rate of 90%, three to six strong seedlings in each plant.

CONCLUSIONThe disinfection method suitable for tiller buds and the medium combination suitable for induction, propagation and rooting of adventitious buds are screened out to establish the rapid cultivation system for tiller buds of E. wushanense.

Epimedium ; drug effects ; growth & development ; Plant Growth Regulators ; pharmacology ; Plant Roots ; drug effects ; growth & development ; Plant Shoots ; drug effects ; growth & development ; Plant Stems ; drug effects ; growth & development ; Time Factors ; Tissue Culture Techniques ; methods

Fibrolase is a non-hemorrhagic zinc metalloproteinase isolated from southern copperhead snake venom (Agkistrodon contortrix contortrix) and is capable of degrading fibrin clots resulting from purified fibrinogen or from blood plasma. Alfimeprase, a truncated form of fibrolase, as a clinical agent was successfully completed PhaseII clinical trials.The cDNA of alfimeprase was amplified by recursive PCR, digested with BamHI and HindII, and cloned into pET43.1a, pMALp2X and pMALc2X vectors to generate fusions with NusA, MBP and sMBP(with signal peptide), respectively. Nus/alfimeprase was expressed in soluble form by co-expressing with chaperone FkpA and inducing with1mmol/L IPTG. The fusion protein accounted for about 25 % of total protein following cell lysis. Alfimeprase was successfully purifiesd by Ni-NTA affinity chromatography and cleaved by enterokinase. The results demonstrate the fibrinolytic activity of recombinant alfimeprase using fibrin plate assays and fibrinogen hydrolysis.

Objective To explore the relationship of the ratio of plasma adrenomedullin(AM)and endothelin-1(ET-1)with serum concentration of neuron-specific enolase(NSE)in full-term neonates with hypoxic-ischemic encephalopathy(HIE).Methods Plasma concentrations of AM,ET-1 and serum NSE from 32 full-term neonates with HIE were detected by radioimmunoassay(RIA)on the 1,3 and 7 d after parturition,30 neonates in the corresponding periods in our hospital were employed as controls.The infants with HIE were divided into mild,moderate or severe group in terms of diagnostic standard of HIE.Results 1.Plasma concentrations of AM and ET-1 in newborns with mild,moderate or severe HIE were significantly higher than that of control group at 1 d after life with a decline from 3-7 d(Pa

Objective To analyze the correlation factors between CT imaging features of pulmonary embolism(PE)and clinical severity stratification,to explore the value of CT pulmonary angiography (CTPA)in acute PE severity stratification.Methods According to the clinical severity,48 patients with acute PE proved by CTPA were classified into two groups,including 21 critical and 27 non-critical patients. Embolism index,ratio of central pulmonary involvement,ratio of right ventricle maximum minor axis (RVMMA)to left ventricle maximum minor axis(LVMMA),namely RV:LV,dilation of main pulmonary and/or right pulmonary trunk,and dilation of bronchial arteries in both groups were analyzed comparatively. The correlation factors between CT imaging features and PE clinical severity stratification were explored.The correlation between RV:LV and embolism index of 48 patients was analyzed.Results Pulmonary embolism index(22.0%—85.0%,median 38.0%),ratio of central pulmonary involvement(42.5%),RV:LV (0.90—1.90,median 1.30),dilation of pulmonary artery(14 cases),and dilation of bronchial artery (8 cases)in critical group(21 cases)were higher than those corresponding factors(5%—48%,median 21.5%,31.25%,0.80—1.40,median 1.00,5 eases,and 3 eases)in non-critical group(27 cases) (Z=4.27,X~2=5.40,Z=2.58,X~2=11.45,X~2=4.87,P

Objective To introduce the technique and experience in reconstruction of posterior crueiate ligament-posterolateral comer(PCL-PLC)injuries with only one allograft of Achilles tendon. Methods The instable knees in 12 cases with PCL injury combined with three degree chronic PLC injury were treated with PCL reconstruction under arthroscope and PLC reconstruction through posterolateral arc incision.Single bundle grafts of PCL reconstructions in tibial and femoral tunnels were fixed by resorption screws.Fibular collateral ligament(FCL)and popliteofibular ligament(PFL)were reconstructed with reforming Larson(?)method.All reconstruction grafts only needed one Achilles tendon as donator.Total op- eration time was 130 minutes including 90 minutes of PCL reconstruction and 40 minutes of PLC recon- struction.Gradual weight loading was allowed after six weeks of bracing.Results Follow-up for mean 12 months(5-24 months)indicated that tibial“step off”reduction was 83%(10/12)and posterior drawer test of 0-1~+ 75%(9/12).Dial sign evaluated that normal external rotation angle was 75%(8/12).Nor- real varus stress test at 30?knee flexion accounted for 83%(10/12).Scores of Lysholm,Tegner and HSS were 90.5,5.1 and 84.5,respectively(P＜0.01=.Conclusion Synchronized reconstruction of PCL and PLC injuries with only one Achilles tendon can obtain satisfactory clinical result,with less expense and shorter operation time.

Objective To increase the transfection of EGFP-N3 plasmids into 293T cells using ultrasound-targeted microbubbles delivery(UTMD) mediated a peptide nucleic acid (PNA) binding nuclear localization signal (NLS).Methods Antibody-targeted microbubbles were used in the experiments which can specifically recognize the SV40T antigen receptor.The SV40T antigen receptors were expressed on the membranes of 293T cells.The PNA containing the NLS were inserted in the EGFP-N3 plasmid DNA,which increased nuclear localization.Ultrasound-targeted microbubble delivery (UTMD) and the PNA binding NLS were utilized to improve the cytoplasmic import of plasmids and the nuclear intake of the plasmid from the cytoplasm,respectively.The study was divided into five groups:Contrast (group A),Common microbubble + DNA (group B),Antibody-targeted microbubbles + DNA (group C),Common microbubbles + NLS-PNA-DNA (group D),Antibody-targeted microbubbles + NLS-PNA-DNA (group E).Fluorescence microscope was used to observe the fluorescent light in each group;flow cytometry to test the transfection;RT-PCR and Western blot to detect genes' mRNA and protein expression level.Results Ultrasound and antibody-targeted microbubble delivery (UTMD) significantly enhanced the cytoplasmic intake of exogenous genes and maintained high cell viability(＞80％).Fluorescent microscope showed that the quantities of green fluorescence in cells were increased successfully.The transfection results of flow cytometry were 0,(9.30 ± 0.46)％,(26.46 ± 2.01)％,(29.54 ± 0.62)％,(45.72 ± 1.86)％,respectively,and the differences were statistically significant(P ＜0.05).The relative mRNA and protein expression in group E were greater than those in group C and D respectively (P ＜0.05).Conclusions UTMD combined with antibody-targeted microbubbles and a PNA binding NLS plasmid can significantly improve transfection efficiency of exogenous genes by enhancing both cytoplasmic and nuclear DNA import.

Adolescent ; Adult ; Aged ; Cerebrospinal Fluid Rhinorrhea ; etiology ; surgery ; Child ; Endoscopy ; adverse effects ; Humans ; Male ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; adverse effects ; Paranasal Sinuses ; surgery ; Postoperative Complications ; surgery ; Young Adult