Objective To explore the effects of surface functionalized multi-walled carbon nanotubes (FMWCNTs) on the cytotoxicity of human peripheral blood mononuclear cell (PBMC).Methods Five different types of MWCNTs (hydroxylated,carboxylated,aminated,nickel-plated and pristine MWCNTs (P-MWCNTs)) with the same diameter and length were evaluated the dispersion and characterizations in physiological salt solution by transmission electron microscopy.PBMC were isolated by density gradient centrifugation from human peripheral blood,and 5 types of MWCNTs were ultrasonically dispersed in serum-containing medium respectively.After incubation with PBMC for 12,24,48 or 72 h,cytotoxicity was detected by CCK-8 kits.Results All the MWCNTs had well dispersion,especially the F-MWCNTs.Cytotoxicity results showed that all types of MWCNTs could induced PBMC death,and presented dose-dependence manner and a certain degree of time-dependence manner.Compared with the P-MWCNTs,F-MWCNTs changed cytotoxicity statistically,with the hydroxylated,carboxylated,aminated MWCNTs weakened,aminated MWCNTs significant (P＜0.05),nevertheless the nickel-plated MWCNTs increased.Compared with the P-MWCNTs (25 μg/ml),cell viability of PBMC after 24 and 48 h incubation with the same dose of nickelplated MWCNTs both decreased,and the differences was statistically significant (P＜0.01,P＜0.05).Conclusions The functional group modification affects not only the MWCNTs dispersion in medium,but also the cytotoxicity of the MWCNTs on PBMC.

Objective To evaluate and compare the histamine-releasing,potencies of cis-atracurium and atracurium during induction of general anesthesia.Methods Forty-five ASA Ⅰ or Ⅱ patients aged 16-71 yr undergoing elective surgery under general anesthesia were randomly divided into 3 groups (n=15 each):group Ⅰcis-atracurium (stored at 4-8℃)(group CIS1);groupⅡcis-atracurium (stored at room temperature)(group CIS2) and group Ⅲ atracurium (stored at 4-8℃)(group ATR).Anesthesia Was induced with TCI of propofol (Cp 3 μg/ml) and remifentanil (Ce 3-5 ng/ml).A bolus of cis-atracurium 0.15 mg/kg or atracurium 0.75 mg/kg Was given iv over 5-10 s as soon as the patients lost consciousness.Neuro-muscular block was monitored with TOF-Watch(R) SX(Organon,the Netherlands).Single stimulation (0.1 Hz) was apphed to the ulna nerve at wrist.The maximal degree of N-M block,onset time,duration of action and recovery index were recorded.The patients were intubated and mechanically ventilated when N-M block reached the maximal degree.The intubation condition Was evaluated.MAP and HR were continuously monitored.Changes in skin were scored (0=no change,Ⅰ=flushed>120 s,Ⅱ=erytbema,Ⅲ=urticaria).Blood samples were obtained before (T0,baseline),at 2 min after induction of anesthesia with TCI of propofol and remifentanil (T1) and 2 and 5 min after CIS/ATR administration (T2,T3) for determination of plasma histamine concentration using enzymatically amplified immunoassay.Results The onset time was significantly longer and the duration of action was significantly shorter in group CIS1 than in group ATR.The maximal degree of N-M block was 100%and the intubation condition was excellent in group CIS1 and ATR.There wag no significant difference in the recovery index between group CIS1 and ATR.The onset time was significantly longer and duration of action shorter in group CIS2 than in group CIS1.There was no significant difference in recovery index between group CIS1 and CIS2.There was no significant change in plasma histamine concentration at T1-3 as compared with the baseline at T0 in group CIS1 but plasma histamine concentration was significantly increased at T2,3 in group ATR.MAP was significantly decreased after induction of anesthesia with propofol and remifentanil,but CIS and ATR did not significantly change MAP.Conclusion The onset time is longer and duration of action is shorter after cis-atracurium than afar atracurium.The N-M block induced by cis-atracurium is significantly attenuated if stored at the room temperature.Cis-atracurium does not cause histamine release.

Objective To investigate familial hyperlipoproteinemia and the features of familial hyperlipoproteinemia in Lanzhou . Methods Data were from previous studies on the subject .Families of hyperlipoproteinemia were screening ,questionnaires were col-lected ,physical examination and laboratory data of family members were also colleted to analysis the characteristics of familial hy-perlipoproteinemia .Results A total of 39 familial hyperlipoproteinemia families were enrolled in the study ,including 280 family members .There were 15 core families ,11 single-parent families ,and 13 orphaned families .There were 6 familial hypercholesterol-emia families ,9 familial hypertriglycerides families ,24 mixed familial hyperlipidemia families .The children of the first generation ac-counted for 63 .2% of the total number of people enrolled in the study ,the father generation accounted for 14 .3% ,the children of the second geration accounted for 22 .5% .Conclusion In the survey ,the most common type of familial hyperlipidemia was mixed familial hyperlipidemia .The father generation was majority .The member of core families was less than incomplete families .

Objective To investigate the effect of early continuous veno-venous hemofiltration (CVVH) on intra-abdominal pressure (IAP) and serum interleukin-6 (IL-6) in severe acute pancreatitis (SAP) patients with abdominal compartment syndrome (ACS).Methods 41 SAP patients with ACS were selected as treatment group and treated with CVVH as well as conventional methods in ICU.The other 12 patients with the same disease were selected as the control group and were only treated with conventional methods because of economic reasons.IAP and blood level of IL-6 in the two groups were measured daily in order to investigate their changes and the correlation between the two parameters.Results The serum IL-6 level and IAP in the two groups were higher on admission day.IAP and serum IL-6 level in the treatment group were significantly decreased on the first day after treatment,and thereafter decreased rapidly.In the control group,IAP and serum IL-6 level were significantly decreased on the 3rd day after treatment.IAP and serum IL-6 level from the 1st day to the 6th day after treatment in the treatment group were significantly lower than those of the control group at the same time point (P＜0.05).There was a significant positive correlation between blood IL-6 level and IAP in SAP patients with ACS(r=0.48,P＜0.01).IL-6 difference before and after treatment was also positively correlated with the difference of IAP(R=0.39,P＜0.05).Conclusions VVH significantly decreased the IAP and the blood level of IL-6 in ACS patients of SAP.The blood level of IL-6 is significantly correlated with IAP,suggesting that IL-6 may play an important role in the pathogenesis of ACS.Therefore early CVVH may clear the cytokines such as IL-6 and lower IAP,thus to prevent multiple organ dysfunction syndrome (MODS),which should be applied in the early stage of ACS.

Objective To prepare stable aqueous dispersions of chitosan/multi-walled carbon nanotubes (CS/MWCNTs) composites,and observe the effects of CS/MWCNTs on the growth of human umbilical vein endothelial cells (HUVEC).Methods CS/MWCNTs composites were prepared by electrostatic interactions between negatively charged MWCNTs and positively charged low-molecular-weight CS.The prepared CS/MWCNTs were characterized by transmission electron microscopy and Zetasizer nano-analyser.The cellular uptake of the fluorescently labeled CS/MWCNTs was observed by laser confocal microscopy after incubating with HUVEC for 24 h at different concentrations.In vitro cytotoxicity and cellular reactive oxygen were also detected.Results When the mass ratio of low-molecular-weight CS to MWCNTs was equal or greater than 10∶1,the CS/MWCNTs can be stabilized in solution.Cellular uptake experiments showed that the CS/MWCNTs could enter into the cells and locate mainly in the cytoplasm.Cytotoxicity study showed that the CS/MWCNTs composites was less toxic than MWCNTs alone at high concentration (10 and 20 μg/ml).However,there was no significant differencein the level of cellular reactive oxygen between the two groups (P＜0.05).Conclusions CS/MWCNTs composites showed low cytotoxicity and high stability,which would be a promising carrier for drug delivery.

ObjectiveTo explore the endocytic pathway of TAT-LHRH modified chitosan/DNA nanoparticle (TLCDN) that exhibits high transfection efficiency and targeting to HepG2.MethodsPlasmid DNA was labeled with fluorescein,and the resulting fluorescent DNA was complexed with chitosan or TAT-LHRH modified chitosan to form chitosan/DNA nanoparticle (CDN) and TLCDN by the complex coacervation method.Internalization of TLCDN or CDN by HepG2 cells were measured in the presence of three kinds of inhibitors of endocytic pathway,Chlorpromazine,Filipin or Dynasore,using High-Content Analyzer to collect and analyze the data.ResultsChlorpromazine led to more decreased uptake of CDN than that of TLCDN,although not statistically significant.Filipin demonstrated significant inhibitory effect on the uptake of TLCDN while promoted the uptake of CDN.Dynasore resulted in a similar decrease in the uptake of both nanoprticles.ConclusionIt was demonstrated that CDN was taken up by HepG2 cells mainly through the clathrin-dependent endocytic pathway and TLCDN was more likely to be internalized by HepG2 cells through the caveolin-mediated endocytic pathway although the clathrin-dependent endocytic pathway was also involved.

Objective To analyze the effect of dexmedetomidine hydrochloride injection on patients with craniocerebral disease who has no artificial airway in the process of bronchoalveolar lavage treatment.Methods Forty-six patients (age 17-28,average age 56.6±9.2,26 men and 20 women) with craniocerebral disease who has no artificial airway were selected,and were treated by bronchoalveolar lavage for lung infection.The patients were randomly divided into two groups,control and test group.The control group (n=23) received midazolam for sedative and the test group (n=23) received dexmedetomidine hydrochloride for sedative while they were in the process of bronchoalveolar lavage treatment.Heart rate,mean arterial pressure and blood oxygen saturation of fingers collected from patients before and during the process of bronchoalveolar lavage were compared.Results In the process of bronchoalveolar lavage treatment,the minimum blood oxygen saturation of finger artery from the control group was lower than that from the test group,the fastest heart rate from the control group was greater than that from the test group,and the lowest mean arterial pressure from the control group was lower than that from the test group (P＜0.05).In two groups,heart rate in the process of bronchoalveolar lavage treatment was faster than that from before the treatment,while both mean arterial pressure and blood oxygen saturation of finger artery were decreased (P＜0.05).Conclusions Continuous intravenous pumping of dexmedetomidine hydrochloride on patients with craniocerebral disease who has no artificial airway during the process of bronchoalveolar lavage treatment is effective and safe,and it has less inhibitory effect on respiratory function and blood pressure.

Objective To investigate the impact of chitosan and alkylated chitosan DNA nanoparticles on the function of human naive CD4+T cells.Methods The secretion of cytokines (IL-4 and TNF-γ) was observed after the co-incubation of human naive CD4+T cells with nanoparticles 12 h,24 h and 48 h,respectively.ResultsNone of the nanoparticles induced the production of cytokines ( IL-4 and TNF-γ ).Conclusion Chitosan and alkylated chitosan DNA nanoparticles will not induce the differentiation of human naive CD4+ T cells into T1 or T2 and may be considered as a safe gene carrier.

Objective To generate recombinant Pichia pastoris for high-copy expression of human tissue factor pathway inhibitor (TFPI). Methods The cDNA encoding human TFPI was inserted into the expression vector pPIC9K and the constructed expression vector rhTFPI-pPIC9K was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant plasmids were subsequently transformed into Pichia pastoris GS115 cells, and the transformants were confirmed by PCR amplification of the genomic DNA.The recombinant Pichia pastoris with high copies of TFPI cDNA was screened by G418 selection. Western blot and TFPI ELISA Kit were employed to analyzing. The temperature, time and concentration of methanol for the induction of recombinant protein were optimized. Results PCR analysis and DNA sequencing confirmed the successful construction of the expression vector rhTFPI-pPIC9K. Real time quantitative PCR and Western blot analysis demonstrated the positive correlation between TFPI expression level and the copy number of TFPI cDNA in Pichia pastoris cells. Optimization of the induction condition significantly elevated the expression level and activity of TFPI (9.95±0.78 mg/L and 3.91±1.37 U/mL). Conclusion The Pichia pastoris strain with high copy of TFPI expression was successfully constructed, which lays a solid foundation for the further investigation on the function of TFPI and its application in the prevention and therapy of diseases.