Specific scAbs could be obtained through biopanning from phage antibody libraries by use of antigens as target molecules. scAbs specific to thrombin were separated from mouse antibody library by the panning strategy of alternating liquid-solid phase in this paper. Thrombin was biotinylated by photobiotin at first, then avidin-coated magnetic beads were utilized to isolate specific scAbs. The eluted phages were amplified and subject to the second round panning in microtiter plate to remove the unspecific reombinant phages. 4 specific scAbs were separated from 23 phage clones after four rounds of alternative panning.

Child, Preschool ; Humans ; Infection ; complications ; Jugular Veins ; pathology ; Male ; Pharynx ; Rupture ; etiology

BACKGROUND:It remains controversial about the clinical outcomes of bone fil ing mesh containers (BFMCs) and percutaneous kyphoplasty (PKP) in pain relief, kyphosis correction, vertebral height restoration and reduction of cement leakage. OBJECTIVE:To compare the clinical outcomes of BFMCs and PKP for osteoporotic vertebral compressive fracture. METHODS:A total of 90 patients with osteoporotic vertebral compressive fracture were equivalently randomized into two groups, fol owed by treated with BFMCs or PKP, respectively. During a more than 3-month fol ow-up, pain relief, kyphotic angle, the vertebral height and cement leakage were observed in the two groups to assess the therapeutic effects. RESULTS AND CONCLUSION:Pain in al patients was relieved at 24 hours after operation. There was no significant difference in pain relief between two groups (P>0.05). PKP was more effective to restore the vertebral height (P<0.05), while BMCFs significantly reduced the leakage rate of bone cement (P<0.05). These results suggest that BFMCs and PKP have their own advantages in the treatment of osteoporotic vertebral compressive fracture, but both exert analgesic effects.

Objective To evaluate clinical effectiveness of transcatheter closure of atrial septal defects ( ASD) with severe pulmonary arterial hyperyension ( sPAH) by fenestrated Amplatzer septal occluders ( ASO) . Methods From September 2002 to April 2013, 17 patients of ASD with sPAH received transcatheter ASD closure using fenestrated occluders. Aged from 18 - 72 years, the diameters of ASDs were 18 - 33 mm. The systolic pulmonary arterial hypertension measured by transthoracic echocardiogram were 80 - 112 (96. 9 ± 8. 9) mmHg. The follow-up study included electrocardiography, chest radiography and echocardiography. All the patients were followed up for 1. 5 - 12 ( mean 6. 4 ± 0. 7) years. Results Systolic pulmonary arterial pressure (sPAP) of 60 - 108 (88. 7 ± 11. 7) mmHg and mean pulmonary artery pressure ( mPAP) of 29. 3 - 60 (51. 0 ± 8. 1) mmHg were measured by cardiac catheterization before ASD closure. Qp/ Qs was 1. 50 - 2. 44 (1. 8 ± 0. 31) and the pulmonary vascular resistance was 3. 1 - 9. 7 (5. 6 ± 1. 5) wood units (wu) . Immediately after the implantation of fenestrated occluders, sPAP decreased to 56 - 99 (70 ± 11. 5) mmHg and mPAP to 27 - 51. 7 (41. 1 ± 7. 1) mmHg. On the 3 d, 3 m and 6 m follow-up exam, RVEDd decreased ( P ﹤ 0. 05), while LVEDd, LVEDV and LVEF increased significantly (P ﹤ 0. 05) . sPAP decreased significantly after transcatheter closure at 3 m and 6 m as compared to pre-closure levels (both P ﹤ 0. 05) . The mean sPAP in long term follow up was (60. 2 ± 13. 3) mmHg which had significant decrease compared to pre-closure level ( P ﹤ 0. 01), but no significant difference found when compared to 6 m follow up (P ﹥ 0. 05). Conclusions ASD closure with fenestrated ASO is a satisfactory approach for ASD with severe PAH.

To investigate the protective effect of the seed oil of Abelmoschus esculentus on gastric ulcer,two acute gastric ulcer mice models were established by intragastric administration of aspirin or absolute ethanol,respectively.Clinical index of ulcer area,ulcer index,gastric volume,gastric pH value,free acidity,total acidity,and histopathological assessment were measured to evaluate the injuries of gastric ulcer and the protective effect of okra seed oil In order to comprehensively uncover the possible underlying mechanism,a series of biochemical assays were also performed,including serum TNF-α,IL-6,IL-10 and Tbil,NO,MPO and SOD in the stomach included.Moreover,the ALT,AST and ALP in the liver of mice were also tested to evaluate the possible hepatic toxicity of the seed oil.The results indicated that the seed oil of A.esculentus exerted protective effect in ethanol-induced gastric ulcer mice by reducing the ulcer area and ulcer index,declining the free and total acidity,and increasing the pH value of gastric content.Histopathological observation showed the gastric mucosa of the acute gastric ulcer mice induced by alcohol was incomplete and severely damaged,with submucosal edema and nuclear pyknosis,as well as glandular structure disappearing,compared with that of normal mice.What's more,a number of inflammatory cell infiltration occured in the gastric mucosa of alcohol-model mice,with messes of neutrophils,lymphocytes,eosinophils and plasma cells.Okra seed oil could improve the damaged structure of the gastric mucosa and gland caused by ethanol,but could not ameliorate the condensation of nucleus and infiltration of inflammatory cells.Biochemical analysis revealed that the seed oil of A.esculentus could counteract the damage induced by ethanol via decreasing Tbil and TNF-α in serum,decreasing NO and myeloperoxidase,and increasing SOD in stomach.Meanwhile,okra seed oil exhibited protective effect in aspirin-induced gastric ulcer mice by increasing the gastric content pH,and reducing free and total acidity.Compared with the control group,the gastric mucosa of aspirin-model group showed multifocal coagulation necrosis,sheet edema and infiltration of inflammatory cells by histopathological assessment.Compared with the aspirin-model group,the soybean oil group and okra seed oil group could ameliorate the inflammatory cell infiltration.Biochemical analysis revealed that okra seed oil could counteract the injury induced by aspirin via decreasing TNF-α and IL-6,and increasing IL-1O in serum,decreasing NO and MPO and increasing SOD in stomach.In a word,the okra seed oil exerted protective effect on acute gastric ulcer by anti-inflammation,anti-oxidation and hepatocyte protection.The okra seed oil deserves further development and utilization.

Alanine Transaminase ; blood ; Animals ; Bilirubin ; blood ; Disease Models, Animal ; Endotoxemia ; etiology ; Endotoxins ; blood ; Gastrointestinal Hormones ; blood ; Gastrointestinal Motility ; physiology ; Intestine, Small ; physiopathology ; Liver ; pathology ; Liver Failure, Acute ; blood ; etiology ; physiopathology ; Liver Function Tests ; Rats ; Thioacetamide ; administration & dosage

OBJECTIVETo study the changes of partial pressure of brain tissue oxygen (PbtO2) and brain temperature in acute phase of severe head injury during mild hypothermia therapy and the clinical significance.

METHODSOne hundred and sixteen patients with severe head injury were selected and divided into a mild hypothermia group (n=58), and a control group (n=58) according to odd and even numbers of hospitalization. While mild hypothermia therapy was performed PbtO2 and brain temperature were monitored for 1-7 days (mean=86 hours), simultaneously, the intracranial pressure, rectum temperature, cerebral perfusion pressure, PaO2 and PaCO2 were also monitored. The patients were followed up for 6 months and the prognosis was evaluated with GOS (Glasgow outcome scale).

RESULTSThe mean value of PbtO2 within 24 hour monitoring in the 116 patients was 13.7 mm Hg +/- 4.94 mm Hg, lower than the normal value (16 mm Hg +/- 40 mm Hg ) The time of PbtO2 recovering to the normal value in the mild hypothermia group was shortened by 10 +/- 4.15 hours compared with the control group (P<0.05). The survival rate of the mild hypothermia group was 60.43%, higher than that of the control group (46.55%). After the recovery of the brain temperature, PbtO2 increased with the rise of the brain temperature.

CONCLUSIONSMild hypothermia can improve the survival rate of severe head injury. The technique of monitoring PbtO2 and the brain temperature is safe and reliable, and has important clinical significance in judging disease condition and instructing clinical therapy.

Adult ; Blood Gas Monitoring, Transcutaneous ; Body Temperature ; Brain ; metabolism ; Brain Chemistry ; Craniocerebral Trauma ; metabolism ; therapy ; Female ; Humans ; Hypothermia, Induced ; Male ; Oxygen ; metabolism ; Partial Pressure ; Regional Blood Flow ; Treatment Outcome

Bacterial Proteins ; chemistry ; Bacterial Secretion Systems ; Crystallography, X-Ray ; Legionella pneumophila ; metabolism ; Models, Molecular ; Protein Structure, Tertiary

Objective To study the dynamic changes of cytokines including tumor necrosis factor-alpha (TNF-?),interleukin-6 (IL-6),IL-8 in serum and cerebrospinal fluid (CSF) of asphyxia neonates,and to analyze the relationship between cytokines levels and severity of brain damage and neurological outcome. Methods The concentrations of TNF-?,IL-6,IL-8 in serum and CSF were measured by radioimmunoassay in 63 asphyxia neonates. Neurological development was evaluated at 12 months by children′s developmental scale of china.Results The serum concentrations of TNF-?, IL-6,IL-8 were significantly higher in asphyxiated neonates than those in the controls,and they were correlated with the degree of encephalopathy. The level of serum TNF-? was hig-(hest) at the first day and IL-6 was highest at the third day. There was no marked dynamic changes within 5 days in serum IL-8 level. The concentrations of TNF-?,IL-8 in CSF were higher at the first and the third day.The dynamic changes of IL-6 in CSF were similar in serum and they were positively correlated. The serum concentration of IL-6 in severe brain injury group was much higher than those of normal and mild group.The CSF concentration of IL-6 in severe brain injury group was much higher than that of normal group. The CSF concentration of IL-8 in severe brain injury group was much higher than those of the normal and mild group. Conclusions The concentrations of TNF-?,IL-6 and IL-8 are increased both in serum and CSF in asphyxiated neonates which are correlated with severity of hypoxic-ischemic encephalopathy. Cytokine-mediated inflammatory reactions may participate in the mechanism of hypoxic-ischemic brain injury after asphyxiaion.The concentration of IL-6 in serum and IL-6, IL-8 in CSF are correlated with the neurological outcome.

BACKGROUNDCisplatin (DDP) is one of most effective and most commonly used therapeutic agent in treating tumors, it can accumulate in the kidney and lead to acute renal failure. MicroRNA-181a can induce cell apoptosis by suppressing the expression of Bcl-2 family. In the present study, we investigated the role of microRNA-181a in the apoptosis of tubular epithelial cell induced by DDP.

METHODSHK-2 cells were cultured, transfected with microRNA-181a inhibitor for 48 hours, and stimulated with 50 µmol/L cisplatin for 24 hours. MicroRNA-181a expression was analyzed by real time PCR, and cell apoptosis was detected by flow cytometry. Moreover, Bcl-2 and Bcl-2-associated X protein (Bax) expression were measured by Western blotting.

RESULTSMicroRNA-181a expression significantly down-regulated in cells transfected with microRNA-181a inhibitor, compared with that in untransfectd cells (21.19 ± 2.01 vs. 38.87 ± 1.97, P < 0.05). Cell apoptosis induced by DDP significantly decreased in cells transfected with MicroRNA-181a inhibitor. Compared with DDP treated cells alone, Bcl-2 expression strikingly was up-regulated and Bax expression was down-regulated in cells transfected with microRNA-181a inhibitor.

CONCLUSIONOne pathway of DDP induces apoptosis of tubular epithelial cell by suppressing Bcl-2 expression is achieved by regulating the target gene of MicroRNA-181a.

Apoptosis ; drug effects ; genetics ; Blotting, Western ; Cell Line ; Cisplatin ; pharmacology ; Flow Cytometry ; Humans ; MicroRNAs ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; bcl-2-Associated X Protein ; metabolism