Objective To investigate the cytotoxicity of ADM-PBCA-NP on L-02 cells.Methods L-02 cells were cultured in vitro and the LDH activity of supernatant liquid of culture cells was examined.The toxicity of ADM-PBCA-NP,ADM and PBCA-NP to L-02 cells by the MTT assay was also determined,and the haemolysis function of PBCA-NP with different concentrations was detected.Results The cytotoxicity of ADM-PBCA-NP,ADM and blank PBCA nanoparticles to L-02 cells under the 10-6 mol/L concentration range was not cytototic(grade 1).LDH activity of supernatant liquid of culture cells showed no differences between the 3 groups.Conclusions Nanoparticles of ADM-NP and PBCA-NP in the 10-6 mol/L concentration range have no significant toxic effect on L-02 hepatic cells;and in a certain concentration range,the cell compatibility is excellent and will not lead to hemolysis.

Air ; analysis ; Chromatography, Gas ; methods ; Ketones ; analysis ; Workplace

Objective To explore the high risk factors, pathogenesis and methods of early diagnosis of periventricular leukomalacia(PVL) in premature infants.Methods The history of intrauterine hypoxia-ischemia was investigated in premature infants;TORCH-IgM antibodies in cord blood of premature infants were measured by ELISA; Nitric oxide (NO) levels in their cord blood were determined by nitric acid reducing enzyme means. Results Thirty-nine of 52 premature infants in PVL group had a history of intrauterine hypoxia-ischemia; TORCH-IgM antibody positive rate in cord blood of premature infants in PVL group was significantly higher than that in control group(P

BACKGROUND: Bone morphogenetic proteins (BMPs) are involved in the formation of various tissues including bone, cartilage, tendon, and ligament. Vascular endothelial growth factor (VEGF) promotes angiogenesis by increasing the permeability and migration of endothelial cells.OBJECTIVE: To construct a co-expressing vector of human bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor 165 (VEGF165), and observe the expression of BMP2 and VEGF165 in human bone marrow mesenchymal stem cells (hBMSCs).DESIGN: Observation control trail.SETTING: Department of Plastic Surgery, Affiliated Hospital of Luzhou Medical College and Plastic Surgery Hospital of Peking Union Medical College, Chinese Academy of Medical Sciences.MATERIALS: The MSCs derived from the healthy adult volunteers of marrow donors. pIRES-EGFP-hVEGF165 containing total length of cDNA sequence of human VEGF165 gene was provided by Dr. Cheng Ting from Plastic Surgery Hospital of Peking Union Medical College. pSP65-hBMP2 containing total length of cDNA sequence of human BMP2 gene was provided by Dr. Guo Ximin from the Academy of Military Medical Sciences. Enkaryotic expression vector pIRESneo (Clontech Company) Pyrobest DNA Polymerae, restriction enzyme, DNA ligase and plasmid extraction kit, DNA Fragment Purification Kit (TaKaRa Company), LiorfectamineTM liposome transfection kit, DMEM medium, trypase, TRIzoIRNA extraction kit (Gibco BRL), Omniscript RT kit (Qiagen), TaqplusDNA polymerase (Promega), PMSF, leupeptin, aprotinin, chymostatin (Sigma), protease inhibitor, PVDF membrane (Amersham-Pharmacia Biotech), rabbit anti-human BMP2 antibody and VEGF monoclonal antibody (Santa Cruz Company), goat anti-rabbit lgG-peroxydase (Wuhan Boster), G418 (Ameresco Company in U.S).METHODS: The experiment was conducted in the Affiliated Hospital of Luzhou Medical College and the Plastic Surgery Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences between June 2005 and April 2006.hBMP2 and hVEGF165 cDNA were directional cloned into multiple clone sites of the eukaryotic expression vector pIRESneo. The recombinant plasmid was identified by restrictive enzyme Xho Ⅰ/Bgl Ⅱ digestion analysis and DNA sequencing. Liposome-mediated gene transfer method was used to transfect hBMSCs. For observation, the transfected cells were divided into IRES-hBMP2-VEGF165 group, pIRES-hBMP2 group, pIRES-VEGF165 group and empty vector group, which were transfected with pIRES-hBMP2-VEGF165, pIRES-hBMP2, pIRES-VEGF165 and pIRES-neo. Meanwhile, the same number nontransfected cells were selected as blank control group. The reverse transcription polymerase chain reaction (RT-PCR) and Weszern blot were employed to observe the expression and secretion of hBMP2 and hVEGF165 gene and protein.expression vector hBMP2 and hVEGF165 gene sequence were the same as reported after restrictive enzyme EcoRI and Bgl Ⅱ digestion analysis and pIRES-BMP2 gene sequencing, which showed that the recombinant plasmid pIRES-hBMP2-VEGF165 highly expressed hBMP2-mRNA and VEGF165-mRNA, but the non-transfected or transfected with pIRES-hBMP2-VEGF165 or pIRES-hBMP2 secreted a great quantity of hBMP2, but that non-transfected or transfected with pIRES-VEGF165 or empty vector secreted only little.CONCLUSION: The co-expressing vector of hBMP2 and hVEGF165 can be expressed stably in hBMSCs.

Acrosin ; immunology ; isolation & purification ; Humans ; Male ; Membrane Proteins ; immunology ; isolation & purification ; Protein Binding ; Spermatozoa ; metabolism ; Zona Pellucida ; metabolism

OBJECTIVETo establish a rapid gas chromatographic method for determination of o-chlorostyrene in the air of workplace.

METHODThe air samples were collected by syringes, injected directly to the GC system, and then separated by a FFAP capillary column (30 m × 0.53 mm × 0.25 µm), finally determined by a Flame Ionization Detector.

RESULTSA good linear correlation was showed within a range of 0 ∼ 1200 µg/L, with regression formula Y = 14 030 + 7 207X (r = 0.9999). The air sample could be stably stored in the syringe for 5 hrs. The relative standard deviation (RSD) of repeated injection of o-chlorostyrene standard solutions at three different concentration by six times was 1.28% ∼ 1.97%. The minimum detectable concentration was calculated to be 5.2 mg/m(3). Other coexistent violative organic compounds such as styrene, p-chlorostyrene, and m-chlorostyrene didn't interfere with the determination under the experimental conditions of this method.

CONCLUSIONThis method meets the requirement of "Guide for establishing occupational health standards-Part 4: Determination methods of air chemicals in workplace". It is applicable for determination of o-chlorostyrene in the air of workplace.

Objective To explore the application of B-mode intra- and post-operative ultrasound in patients with traumatic brain injuries. Methods B-mode intra- and post-operative ultrasound were performed 35 times in 24 patients, including 7 times of intra-operation in 7 patients and 28 times of post-operation in 17 patients. Results During operation epidural hematomas in 3 cases as well as intracranial hematomas in 2 cases, subdural hematomas in 1 case, diffused brain swelling in 1 case were found. After operation epidural hematomas were examined in 4 cases as well as subdural hematomas in 1 case, delayed intracranial hematomas in 6 cases, scattered contusion and laceration of the brain in 12 cases, acute obstructive hydrocephalus in 1 case, and subdural hydroma in 4 cases. Six cases underwent operations again because of delayed hematomas, contusion and laceration of the brain. B-mode ultrasound was compared with CT scan and the total coincidence rate was 85.4%.Conclusions B-mode ultrasound is helpful to examine delayed hematomas and other emergent complications. It may save time and may improve rescue rate and has a good coincidence with CT scan. B-mode ultrasound may be widely used as a routine way in neurotrauma ICU.

Objective To investigate the role and intracellular signal transduction mechanism in the injury of renal tubular cells induced by postasphyxial-serum in neonate.Methods Human renal proximal tubular cell(HK-2 cell) was used as target cell. The experiment was designed as:control group, asphyxia group ,and pyrrolidine dithiocarbamate (PDTC)blocking group. The attacking concentration of serum was 20%, and the apoptosis rate of HK-2 cells was detected by flow cytometer.Results Compared with controls[(13.3?1.70)%],after being stimulated with postasphyxial-serum, the apoptosis rate of HK-2 cells of asphyxia group [(46.73?3.68)%] and PDTC blocking group [(31.19?2.79)%]were significantly increased(P

Objective To investigate the role of postasphyxial-serum of neonate in inducing injury of human renal proximal tubular cells(HK-2 cells).Methods HK-2 cells were used as target cell.The neonatal different concentration postasphyxial-serum of 1,3,7 days after asphyxia were used as attacking means.The experimental groups were divided into 15 groups:the 2.5%,5.0%,10.0%,(20.0%) attacking concertration groups of 1,3,7 day after asphyxia and control group of each concertration.The culture medium and concertration of the control group and the experimental groups were the same.The changes of morphology were observed under inverted microscope,the cell viability was measured by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the changes in morphology of HK-2 were most serious and obvious,the cell viability were obviously decreased(all P

Objective To investigate the role of erythropoietin(EPO)in relieving the injury of human renal tubular cells (HK-2) induced by postasphyxial-serum of neonates.Methods Human renal proximal tubular cell(HK-2) was used as the target cell.The experiment was designed as control group, asphyxia group,and group of pretreatment with EPO. The attacking concentration of serum was 200 mL/L,then the changes of morphology were observed under inverted microscope,and the cell viability was measured by 3-(4,5-dimethy lthiazcl-2-yl)-2,5-diphenyl-tetazolium bromide(MTT) methods,and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the change in morphology of HK-2 was most serious and obvious,and the leakage rate of LDH increased significantly,and the cell viability decreased obviously in asphyxia group.But compared with asphyxia group,the change in morphology of HK-2 was obviously improved,and the leakage rate of LDH decreased and the cell viability increased in group of pretreatment with EPO in a dose-dependent manner except the group of 1 IU/mL.Conclusion EPO can play the role in relieving the injury of renal tubular cells induced by postasphyxial-serum in neonates.