Objective To investigate the effects of low dose of extracellular histone on endothelial cells in infectious diseases such as sepsis. Methods The endothelial cells were treated with 10 μg/mL recombinant human histone H3/H4 complex in replacement of calf thymus histones (CTH) for various periods of time, and the morphology changes and the viability of the endothelial cells were recorded. In addition, flow cytometry was applied to identify the characteristics of endothelial cells and enzyme-linked immunosorbnent assay (ELISA) was used to detect the extracellular histones level in endothelial cells culture. Results The low dose of CTH could continuously induce endothelial cells death, cell morphological changes and function loss, which was reproduced by 10 μg/mL recombinant histone H3/H4 complex. Results of histones quantitation showed that histone can cause a series of intracellular reactions in a short period of time. Conclusions It is showed that 10 μg/mL H3/H4 can induce the toxicity in infectious disease and this level of the dose is a lower than those used in previous studies and more close to the pathological conditions.

ObjectiveTo investigate the effect of clinical pathway teaching methord in nursing practice teaching. Methods80 junior college nursing students were randomly divided into control and experimental groups. Traditional clinical teaching method was given to control group, while the clinical pathway teaching method was given to observation group. Scores of comprehensive quality after departmental rotation and satisfaction rates of nursing students to teaching method in these two groups were evaluated. ResultsThe experimental group was significantly better than the control group ( P＜0.05 ), and the difference was statistically significant. ConclusionThe clinical pathway can significantly improve the quality of nursing practice teaching.

Objective To study the correlation between blood glucose variability and TOAST type of patients with acute ischemic stroke and provide new evidence which can improve the prognosis of ischemic stroke.Methods One hundred and forty-three hospitalized patients with acute ischemic stroke from August 2013 to August 2014 were included.All patients were estimated about the general state of health and performed laboratory examinations and other auxiliary examinations.Then the TOAST type of all cases was classified.The 24-hour blood glucose monitoring was done to the patients to calculate the blood glucose variability.One hundred and forty-three cases were divided into blood glucose variability group ( standard deviation of glucose ( GluSD )≥1.4 mmol/L, 85 cases ) and control group ( GluSD <1.4 mmol/L, 58 cases) according to glucose variability.We estimated the neurologic impairment of the patients with the US National Institutes of Health Stroke Scale ( NIHSS) and the activities of daily living with Barthel index.The relation between blood glucose variability and the prognosis of ischemic stroke was analysed with multivariate analysis.Results Differences were significant in age, history of diabetes and NIHSS score between blood glucose variability group and control group.The degree of neurologic impairment of large artery atherosclerosis (LAA) group (moderate and severe neurologic impairment:43/53(81.0%)) and cardio-aortic embolism ( CE) group ( moderate and severe neurologic impairment:15/17 ) was more serious than that in small artery occlusion ( SAO) group ( moderate and severe neurologic impairment:6/71 ( 8.5%) ) , especially in CE group.The differences were significant (χ2 =7.043,P<0.05).Blood glucose variability in patients with LAA was more obviously than that in other patients.NIHSS score and activities of daily living of the patients estimated on admission and after 2 weeks were not different in the blood glucose variability group, but the difference was significant in control group.The poor prognosis in blood glucose variability group was 2.821 times that in control group ( 95% CI 1.880 -4.233 ).Conclusions The people sufferd ischemic stroke with old age, diabetes or severe case are more vulnerable to abnormal blood glucose variability.Abnormal blood glucose variability is an independent risk factor for acute ischemic stroke and induces difficulty to recovery and poor prognosis.

Objective To investigate the change of extracellular histone level as well as the mecha-nism of the cytotoxicity of extracellular histones on vascular endothelial cell in sepsis. Methods Septic chil-dren admitted to PICU in Shanghai Children′s Medical Center between January 2010 and December 2014 were included in the present study. According to the diagnostic criteria of sepsis,the patients were divided into the sepsis group(51 cases) and the severe sepsis group(79 cases),with healthy children as the control group (108 cases). Patients in the severe sepsis group were further divided into the survival group(45 cases) and the non-survival group ( 34 cases ) based on 28-day mortality. The plasma concentration of extracellular histones in these children was determined and its correlation with the severity of sepsis was analyzed. Human umbilical vein endothelial cells(HUVECs) were incubated with calf thymus histone(CTH) at various con-centrations(0,50,100,200 and 300μg/ml) or different time periods(200μg/ml,0,5,15,30,45 and 60 mi-nutes) . The treated cells were subject to flow cytometer to measure the cell survival rate and scanning/trans-mission electron microscopy to observe their morphological changes. Western blot was used to detect the ex-pression of IκB and phosphor-p38/p38 in nuclear factor ( NF )-κB and mitogen-activated protein kinase ( MAPK) signaling pathways,while ELISA was used to determine the levels of tumor necrosis factor-α and interleukin-6. Results The levels of circulating histones in the septic children(2. 29 ± 1. 00) and severe sep-tic children ( 19. 17 ± 10. 20 ) were significantly higher than that of healthy controls ( 0. 23 ± 0. 26 ) ( P <0. 001),and the histone levels in the severe septic children were even higher(P<0. 001). Among the chil-dren diagnosed as severe sepsis,the level of circulating histones in the non-survivors was significantly higher than that in the survivors(29. 47 ± 5. 99 vs. 10. 94 ± 2. 68,P<0. 001). The survival rate of HUVEC gradually decreased along with the increase of CTH concentration or the treatment period in vitro. Data from electron microscopy showed that CTH treatment could directly disrupt the plasma membrane of HUVEC. Histones could also activate NF-κB and MAPK pathways,leading to the release of large amount of tumor necrosis fac-tor-α and interleukin-6. Conclusion The levels of extracellular histones in the septic children are correlated with the severity of sepsis. CTH can induce HUVEC death in a dose-and time-dependent manner. Extracellu-lar histone-induced endothelial dysfunction may mediate the progression of sepsis and such cytotoxicity might be due to the destruction of endothelial cell membranes and activation of the NF-κB and MAPK signaling pathways.

Objective To investigate the effect of local injection of Botulinum toxin type A(BTXA) on spasticity and function of the affected upper limb in stroke patients.Methods A total of 32 stroke patients were re- cruited and randomly divided into two groups:a BTXA group and a control group.All the patients had spasticity of upper limb muscles,which scored grade 2 to 3 with the Modified Ashworth Scale(MAS) ,and decreased elbow joint range of motion.The 16 patients in the BTXA group received BTXA injection in the biceps brachii muscles and flexor muscles of forearm on 10～15 points,while those in the control group did not.All the patients in both groups were treated with rehabilitation training techniques.The MAS,Fugl-Meyer upper limb function assessment and Barthel In- dex were employed to evaluate the changes of muscle tone,upper limb function and activity of living (ADL)perform- ance of the patients before injection and at 1st,2nd,6th 12th weeks after injection.Results The therapeutic effect between the BTXA group anti control group was significantly different in terms of biceps muscle tone,the scores of Fugl-Meyer upper limb function assessment and Barthel Index.Compared with preinjection,muscle tone was de- creased significantly and ADL performance was improved after injection in BTXA group.The effects of BTXA lasted more than 12 weeks.Conclusion Intramuscular muhipoint injection of BTXA was useful in reducing muscle spas- ticity,and was helpful for increasing motor ability of the affected upper limb and ADL performance of the stroke pa- tients.

OBJECTIVETo explore the clinical effects of anterior decompression and fusion with a nano-hydroxyapatite/ polyamide 66 (n-HA/PA66) cage in treating lower cervical fracture and dislocation.

METHODSFrom January 2008 to December 2010, the clinical data of 42 patients with lower cervical fracture and dislocation were retrospectively analyzed. There were 29 males and 13 females aged from 20 to 65 years old. The mean age was 46.8 years. Five cases got injuried in C3, 14 cases in C4, 12 cases in C5, 7 cases in C6 and 4 cases in C7. According to Frankel grade, 4 cases were classified in grade A, 11 cases in grade B, 13 cases in grade C, 9 cases in grade D and 5 cases in grade F. Twenty-eight cases were treated with anterior corpectomy and fusion and 14 cases with anterior discectomy and fusion. Frankel grade was used to do neurologic assessment and visual analogue scale (VAS) was used to evaluate the improvement of clinical symptoms. Segmental height and sagittal lordosis were measured by radiographs and cage location. Cage appearance and fusion status were assessed by 3D-CT images.

RESULTSAll patients were followed up for 3 to 5.2 years with an average of 4.1 years. Frankel grade had obviously improved than preoperative (Z = -4.845, P < 0.001). There were 2, 3, 11, 8, 11 cases classified in grade A, grade B, grade C, grade D and grade E respectively. At the third day after operation and latest follow-up,VAS was (2.6 +/- 1.8),(1.3 +/- 1.0) scores respectively. Both had improved than preoperative (P < 0.05). Up to the latest follow-up, there was only one patient (2.4%) with slight cage translocation (less than 2 mm), however, no cage prolapsed, or collapse, or breakage were found. Both segmental height and lordosis improved significantly after surgery (P < 0.001). And there was not significant difference in both parameters between each postoperative time points (P > 0.05). The mean distance of cage subsidence was 1.5 mm and the rate of cage subsidence (> 3 mm) was 4.8%.

CONCLUSIONThe n-HA/PA66 cage can not only restore and maintain the fusion segmental height and radian, but also promote the osseous fusion and profit the radiographic assessment after operation. Thus, it was an ideal material for prop graft.

Adult ; Aged ; Cervical Vertebrae ; injuries ; surgery ; Decompression, Surgical ; methods ; Female ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fractures ; surgery ; Spinal Fusion ; methods

The purpose of this study is to develop glipizide push-pull osmotic pump (PPOP) tablets by using a formulation design expert system and an artificial neural network (ANN). Firstly, the expert system for the formulation design of osmotic pump of poor water-soluble drug was employed to design the formulation of glipizide PPOP, taking the dissolution test results of Glucotrol XL as the goal. Then glipizide PPOP was prepared according to the designed formulations and the in vitro dissolution was carried out. And in vivo evaluation was carried out between the samples which were similar to Glucotrol XL and the Glucotrol XL in Beagle dogs. The range of the factors of formulation and procedure, which could influence the drug release, was optimized using artificial neural network. Finally, the design space was found. It was found that the target formulation which was similar to Glucotrol XL in dissolution test could be obtained in a short period by using the expert system. The samples which were similar to Glucotrol XL were bio-equivalent to the Glucotrol XL in Beagle dogs. The design space of the key parameter coating weight gain was 9.5%-12.0%. It could be concluded that a well controlled product of glipizide PPOP was developed since the dissolution test standard of our product was more strict than that of Glucotrol XL.
Animals ; Area Under Curve ; Delayed-Action Preparations ; Dogs ; Drug Compounding ; methods ; Drug Delivery Systems ; Drug Design ; Expert Systems ; Female ; Glipizide ; administration & dosage ; chemistry ; pharmacokinetics ; Hypoglycemic Agents ; administration & dosage ; chemistry ; pharmacokinetics ; Male ; Neural Networks (Computer) ; Osmosis ; Polyethylene Glycols ; chemistry ; Random Allocation ; Sodium Chloride ; chemistry ; Solubility ; Tablets

OBJECTIVETo give some theory support of Cistanche tubulosa cultivation by searching dry matter accumulation and echinacoside content of C. tubulosa.

METHODDry matter accumulation content of C. tubulosa culturing in Huabei plain was analysed in different growth season of C. tubulosa. Echinacoside content was determined by HPLC.

RESULTDry matter accumulation of C. tubulosa showed "S" variation. Dry matter accumulation increased fastest in September among growing seasons. Dry matter amount was 138.58 g after C. tubulosa grew a year. Dry matter amount decreased significantly along with inoculation time retarded. Echinacoside content was 30.59% when C. tubulosa grew in 5 months, decreased guadully after that, and 9.76% in annual.

CONCLUSIONVariation rule of dry matter accumulation and echinacoside content was found in C. tubulosa that grew one year in Huabei plain.

Biomass ; China ; Cistanche ; chemistry ; growth & development ; Glycosides ; metabolism ; Plants, Medicinal ; chemistry ; growth & development ; Seasons

OBJECTIVETo study risk factors for severe hand, foot and mouth disease (HFMD) complicated by heart and lung failure and treatment experience.

METHODSA total of 198 children with severe HFMD between March and August in 2011 were enrolled. Univariate analysis and logistic regression model were used to analyze the risk factors severe HFMD complicated by heart and lung failure. The effects of combination therapy with immunoglobulin+dexamethasone+ribavirin were observed.

RESULTSUnivariate analysis indicated that HFMD patients with heart and lung failure had higher proportions of consciousness, tachypnoea, abnormal hemodynamics, increased troponin and EV71 infection than HFMD patients without heart and lung failure (P<0.05).Multivariate logistic regression analysis indicated that tachypnoea, abnormal hemodynamics and EV71 infection were the main risk factors for heart and lung failure. Compared with combination therapy with dexamethasone+ribavirin, combination therapy with immunoglobulin+dexamethasone+ribavirin was more effective for preventing hemodynamic changes in children with severe HFMD (P<0.01). Compared with HFMD patients with heart and lung failure, the effect of the combination therapy with immunoglobulin+dexamethasone+ribavirin was better in HFMD patients without heart and lung failure (P<0.01).

CONCLUSIONSThe main risk factors for heart and lung failure in children with severe HFMD include tachypnoea, abnormal hemodynamics and EV71 infection. Early combination therapy with immunoglobulin+dexamethasone+ribavirin can reduce the incidence of heart and lung failure in children with severe HFMD.

Child, Preschool ; Drug Therapy, Combination ; Female ; Hand, Foot and Mouth Disease ; complications ; Heart Failure ; drug therapy ; etiology ; physiopathology ; Humans ; Infant ; Logistic Models ; Male ; Respiratory Insufficiency ; drug therapy ; etiology ; physiopathology ; Risk Factors

AIMTo clone the gene of staphylococcal enterotoxin C2 and express it in the form of a soluble fusion protein in E. coli. Then the activation of SEC2 on mice lymphocyte and its lethal effects on tumor cells were studied.

METHODSStaphylococcus aureus SEC2 gene was cloned into GST gene fusion vector pGEX-4T-1. The resultant plasmid pGEX-4T-SEC2 was used to transform E. coli BL21, where the GST-SEC2 fusion protein was expressed efficiently. The rSEC2 protein was purified with Glutathione Sepharose 4B affinity column and digested with thrombin. The in vitro culture system was utilized to observe the activation of the SEC2 on mice lymphocyte and the lethal effects on tumor cells of the activated mice lymphocyte.

RESULTSThe proper gene of SEC2 was cloned and purified rSEC2 was obtained. The MTT results indicated that rSEC2 have strong ability to stimulate mice lymphocyte to proliferate with a dose-dependent manner. With the proliferation of mice splenic lymphocyte, rSEC2 has a strong lethal effect on tumor cells B16, K562 and K562-AD.

CONCLUSIONIn this study, the gene of SEC2 was cloned and the rSEC2 protein was obtained, which had strong lethal effect on tumor cells B16, K562 and K562-AD.

Animals ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cloning, Molecular ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Female ; Genetic Vectors ; Glutathione Transferase ; genetics ; Lymphocyte Activation ; drug effects ; Lymphocytes ; cytology ; immunology ; Male ; Mice ; Mice, Inbred ICR ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Spleen ; cytology ; Transfection