AIMTo study the protective action of ulinastatin against lipopolysaccharide (LPS)-induced acute lung injury in mice and the mechanism of its action.

METHODSMice were intraperitoneally injected with ulinastatin (50 and 100 ku x kg(-1)) or saline at a period of 12 h, separately, 30 min after the last injection of ulinastatin, except normal control, all mice of other groups were injected a dose of LPS 15 mg x kg(-1) via tail vein. The levels of TNFalpha in serum and lung were measured by ELISA. The expression of TNFalpha mRNA and iNOS mRNA in lung was assayed by RT-PCR. The expression of c-Fos and c-Jun protein in lung was measured by Western blotting method. And the NO2- / NO3- level in serum and MDA in lung were measured with kits.

RESULTSThe levels of NO2- / NO3- and TNFalpha in serum, MDA and TNFa in lung all increased after iv injection of LPS. The expressions of TNFa mRNA, iNOS mRNA, c-Fos and c-Jun in lung of LPS-injected mice were enhanced. Pretreatment with ulinastatin 100 ku x kg(-1) decreased the levels of NO2- / NO3- in serum and lung, reduced the index of lung, and inhibited the expressions of iNOS mRNA and c-Jun in lung induced by LPS in mice, while ulinastatin showed no effect on TNFa level in serum and lung.

CONCLUSIONUlinastatin protected mice from acute lung injury induced by lipopolysaccharides via inhibiting the activation of c-Jun and iNOS mRNA expression.

Animals ; Blotting, Western ; Glycoproteins ; administration & dosage ; pharmacology ; Injections, Intraperitoneal ; Lipopolysaccharides ; Lung ; drug effects ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Nitric Oxide Synthase Type II ; biosynthesis ; genetics ; Protective Agents ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Proto-Oncogene Proteins c-jun ; biosynthesis ; RNA, Messenger ; biosynthesis ; genetics ; Respiratory Distress Syndrome, Adult ; chemically induced ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; biosynthesis ; blood ; genetics

Objective To assess the accuracy of mixed venous oxygen saturation ( S(-v)O2 ) in reflecting the change in CO during off-pump coronary artery bypass grafting (OPCABG) .Methods Twenty-five NYHA Ⅰ -Ⅲ patients of both sexes, aged 50-75 yr, weighing 55-85 kg, undergoing OPCABG, were studied. Anesthesia was induced with midazolam, fentanyl, etomidate and pipecuronium and maintained with propofol infusion and intermittent iv boluses of fentanyl and pipecuronium supplemented with isoflurane if necessary. The patients were mechanically ventilated (VT 8-10 ml/kg, RR 8-10 bpm, I:E 1:2). PETCO2 was maintained at 35-45 mm Hg.Radial artery was cannulated and pulmonary catheter was placed. CI, S(-v)O2 and Hb were monitored and recorded before skin incision, during anastomosis with left anterior descending artery (LAD), right coronary artery (RCA)and left circumflex coronary artery (LCX), when the chest was closed, when the patients' body position was changed and the heart was manipulated. S(-v)O2 and CI were scaled immediately after the pulmonary artery catheter was placed and before anastomosing LAD. Results The CO change in S(-v)O2 was real-time and accurate in reflecting the body positioning and elevation of hearts. There was no simultaneous significant change in CI.Conclusion The CO change in S(-v)O2 is real-time and accurate in reflecting the body positioning and elevation of hearts during OPCABG.

Pluronic modified polyamidoamine (PAMAM) conjugate (PF127-PAMAM) was prepared and the inhibiting effect of MDR against MCF-7/ADR was investigated with doxorubicin (DOX) as model drug. 1H NMR and FTIR spectra showed that the conjugate was synthesized successfully. Element analysis accurately measured that 27.63% amino of per PAMAM was modified by pluronic (PAMAM : PF127, 1 : 35.37 mole ratio). PF127-PAMAM showed an increased size and a reduced zeta potential compared to PAMAM. PF127-PAMAM had lower hemolytic toxicity and cytotoxicity due to the reduced zeta potential and the protection of PF127. Each PF127-PAMAM molecular could load 19.58 DOX molecules, and the complex exhibited sustained and pH-sensitive release behavior. PF127-PAMAM/DOX exhibited weaker cytotoxicity than free DOX in MCF-7 cells; while the complex showed much stronger reverse effect of drug resistance in MCF-7/ADR cells, and resistance reversion index (RRI) was as high as 33.15.
Dendrimers ; pharmacology ; Doxorubicin ; pharmacology ; Humans ; MCF-7 Cells ; drug effects ; Poloxamer ; pharmacology

Objective To develop europium (Ⅲ) [Eu (Ⅲ)] chelated microparticles for homogeneous immunoassay.Methods Anti-human PCT antibodies were labeled with Eu (Ⅲ) chelated nanoscale microparticles as the detection antibody,and another anti-human PCT antibody was labeled with biotin as the solid-phase antibody.Magnetic microspheres labeled with streptavidin were used to separate the complexes of Eu-IgM-PCT-IgM-Biotin.Results In the homogeneous immunoassay,the standard curve fit was not linear.The quadratic curve was Y =19170.12 + 75493.74X-26.00X2(r =0.9986).According to the standard curve,the limit of detection for PCT was 0.04 ng/ml.Conclusion The homogeneous immunoassay which uses Eu (Ⅲ) chelated microparticles is highly sensitive for detection of PCT recombinant antigens and may serve as a promising method to measure serum PCT levels in the future.

OBJECTIVE: To establish a multiplex STR genotyping method for autosomal STR and Y-STR loci in forensic biological practice. METHODS: Widely used autosomal STR loci and Y-STR loci were selected. A set of PCR primers was designed, and a 5-dye fluorescent labeled STR multiplex PCR reagent kit was developed. RESULTS: A kit was developed which can simultaneously detect 15 autosomal STR loci, 10 Y-STR loci, and an Amelogenin. CONCLUSION The 15 autosomal STR plus 10 Y-STR kit in combination with capillary electrophoresis method was used to STR genotyping with accurate and reliable results. The new one-step testing kit can potentially be widely used in forensic cases and DNA databank in the future.
Amelogenin ; Chromosomes, Human, Y/genetics* ; DNA Primers ; Databases, Nucleic Acid ; Forensic Genetics/methods* ; Genotype ; Genotyping Techniques/instrumentation* ; Humans ; Indicators and Reagents ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction

OBJECTIVETo study the effects of GSM 1800 MHz radiofrequency electromagnetic fields (RF EMF) exposure on protein expression profile of human breast cancer cell line (MCF-7), as to exploring the possible effects on normal cell physiological function.

METHODSMCF-7 cells were continuously or intermittently (5 minutes field on followed by 10 minutes off) exposed to RF EMF for different duration (1 hour, 3 hours, 6 hours, 12 hours, or 24 hours) at an average specific absorption rate (SAR) of 3.5 W/kg. The extracted proteins were separated by 2-dimensional electrophoresis and the protein-spot distribution of the silver-stained gels was analyzed by using PDQuest software 7.1. Each experiment was repeated three times.

RESULTSOn the average, around 1100 proteins were detected using pH 4 - 7 IPG strip. There were no differential proteins found under continuous exposure at SAR of 3.5 W/kg for 6 hours. Under other exposure conditions, we found various differentially expressed proteins in exposure groups as compared with the sham-exposed controls. Especially in 3 hours intermittent exposure and 12 hours continuous exposure, eighteen and seven differential proteins were detected, respectively. The categories and functions of these differentially expressed proteins were analyzed by searching of SWISS-PROT protein database, which suggested that these proteins should be related to the functions of biosynthesization, signal transduction, and DNA damage and repair.

CONCLUSIONSData indicated that the protein expression changes induced by RF radiation might depend on exposure duration and mode. Many biological processes might be affected by RF exposure.

Cell Line, Tumor ; radiation effects ; Dose-Response Relationship, Radiation ; Electromagnetic Fields ; adverse effects ; Female ; Gene Expression ; Humans ; Proteome ; Radio Waves

OBJECTIVESTo review the preliminary clinical experience with high-field-strength intraoperative magnetic resonance imaging (iMRI) suite with neuronavigation system in the pituitary adenoma operation with transsphenoidal approach.

METHODSFrom March 2009 to December 2010, 31 patients [range, 29 - 76 years, mean age (47 ± 11) years]of pituitary adenoma were operated with transsphenoidal approach and intraoperatively with a movable 1.5 T high-field-strength iMRI suite in combination with neuronavigation system. Tumor size was 1.8 - 7.3 cm, mean (3.5 ± 1.2) cm. Twenty-five cases were non-functional pituitary adenoma, 4 cases were prolactin-secreting pituitary adenoma, 2 cases were growth hormone-secreting pituitary adenoma. Thirty patients' resection with transnasal transsphenoidal approach were performed, one patient with transoral transsphenoidal approach was performed.

RESULTSIn 12 cases of 30 patients who planed to totally remove tumor, iMRI had revealed residual lesions and resulted in the change of the surgical strategy, 2 invasive cavernous sinus cases no further resection of the tumor because of internal carotid artery encasement, the other 10 cases resected further, eventually. Finally, 8 cases were totally removed. The ratio of total removal tumor was enhanced to 86.7% (26/30) from 60.0% (18/30). There was no perioperative mortality.

CONCLUSIONSHigh-field-strength iMRI suite with neuronavigation system provides valuable information of tumor resection that allows intraoperative modification of the surgical strategy. It could be very helpful to maximize the resection of the pituitary adenoma and minimize the injury to neurological function.

Adenoma ; surgery ; Adult ; Aged ; Cavernous Sinus ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Neuronavigation ; methods ; Pituitary Neoplasms ; surgery

OBJECTIVETo explore the safety and efficacy of the insertion of screws into fused C1-occipital condyle(CC)complex without image guidance in atlantal-cervical nonsegmentation patients.

METHODSThe occipital condyle junction was fixed posteriorly in 10 basilar invagination patients with atlantal-cervical nonsegmentation using polyaxial titanium screws(3.5 mm)inserted unicortically into the CC complex and C2 pedicles,followed by fixation to a 3 mm rod. Drilling was guided by anatomic landmarks. The entry point was at the center of posterior surface of the CC complex. The angle of medicalization was 10-15 degrees. In the sagittal plane,the angle for maximal superior screw angulation was also 10-15 degrees. The screw length to obtain unicortical purchase was 16 to 22 mm. CT scans were obtained before and after the surgery. The length,width,and height of CC complex were measured on computed tomography(CT)preoperatively. The position of screws and the condition of fixation were analyzed on postoperative CT scan. Postoperative complications were recorded. The mean follow-up was(30.2±4.38)months(range: 24-36 months).

RESULTSThe width,length,height of left side CC complex were(7.96±2.23)mm,(16.06±2.73)mm,and(13.76±2.06)mm,and the width,length,height of right side CC complex were(7.84±1.38)mm,(16.66±2.58)mm,and(12.81±2.62)mm. No fracture was identified. There was no screw malposition or neurovascular complication related to screw insertion. No screw loosening or construct failure was observed during the follow-up.

CONCLUSIONSIn patients with atlantal cervical nonsegmentation,the CC complex screws can be safely inserted assisted by microscope without image guidance. Occipital condyle junction fixation using polyaxial CC complex screws is feasible and can be a good alternative where other fixation techniques are not satisfactory.

Bone Screws ; Cervical Vertebrae ; surgery ; Humans ; Microscopy ; Neck ; Spinal Diseases ; surgery ; Spinal Fusion ; methods ; Tomography, X-Ray Computed

Aged ; Beijing ; China ; Emergency Service, Hospital ; Hospitalization ; Humans ; Temperature

This study was aimed to further investigate the function of platelet collagen receptor-glycoprotein VI and to screen its specific inhibitor. The extracellular domain of platelet glycoprotein VI (GPVI) in E. coli was expressed by recombinant technology, the extracellular domain cDNA of GPVI was amplified from pBluescript KS(-)-GPVI plasmid by PCR. Proved by sequencing, the expression vector pET-20b(+)-GPVI was constructed, which was then transformed into E. coli (BL21(DE3)pLysS) and induced by IPTG. The recombinant GPVI was purified on Ni-NTA resin column and renatured in PBS containing GSH and GSSG. The anti-penta His McAb and anti-GPVI polyclonal antibody were used to identify the recombinant GPVI in Western blotting. Collagen binding test was conducted to investigate the biological activity of recombinant GPVI. The results showed that the recombinant GPVI was expressed in E. coli and successfully purified, which was confirmed to be similar to the native GPVI in Western blotting. The recombinant GPVI can bind the type I collagen in dose-dependent manner. In conclusion, the recombinant GPVI can be achieved in E. coli and restore its native characteristics after renaturation.
Blood Platelets ; metabolism ; Blotting, Western ; Escherichia coli ; genetics ; Humans ; Integrin alpha2beta1 ; Platelet Membrane Glycoproteins ; biosynthesis ; genetics ; Protein Binding ; Receptors, Collagen ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; isolation & purification