Objective To prepare monoclonal antibody specifically against carcinoembryonic antigen glypican-3 (GPC3) and its preliminary application. Methods GPC3 was cloned with PCR to pET16b vector and expressed in E. co-liBL21. Spleen cells were obtained from Balb/c mice embedding immunized with purified antigen intraperitoneally, and fused with Sp2/0 cells. Hybridoma cells were screened by indirect ELISA, and identified by Western blot assay using puri-fied protein after the cell fusion. The indirect immunofluorescence method was used to detect the GPC3 expression in HepG2 cell line. Results The prokaryotic expression vector of GPC3 was successfully constructed, and GPC3 was stably expressed in E. coliBL21. A mouse hybridoma cell line secreting monoclonal antibody to GPC3 was obtained. Western blot analysis showed that monoclonal antibody specifically recognized recombinant protein. Monoclonal antibody could be used to detect GPC3 protein expression in HepG2 cell line by indirect immunofluorescence. Conclusion The monoclonal antibody against GPC3 was successfully obtained.

Brain Injuries ; Erythropoietin ; Humans ; Infant, Newborn

The mode of long-chain alkane uptake by Pseudomonas aeruginosa (CGMCC 1.1785) was studied. P. aeruginosa 1.1785 is capable of using solid long-chain alkane as sole carbon source and producing surface active compound as metabolite. The mass transfer limitation in uptake of alkane was confirmed from the observation that interfacial area of eicosane with water dominates the growth rate of this strain. The enhancement of eicosane uptake by rhamnolipid was mainly caused by increase of interfacial area, since the pseudosolubilized alkane can not support the growth of P. aeruginosa 1.1785. Cell surface hydrophobicity was increased dramatically at the initial phase of growth and followed by a gradual decrease, which indicates that different modes are employed by P. aeruginosa 1.1785 at different growth phase. Therefore, the surfactant mediated mode can be negligible in the uptake process, while the directly attachment mode may not work throughout the growth of P. aeruginosa 1.1785. We proposed a novel uptake mode, in which the chemotaxis of this strain plays an important role.

Neonatal brain damage including hypoxia-ischemia encephalopathy(HIE) and intracranial haemorrhage,may cause adverse neurodevelopmental outcome such as cerebral palsy.Earlier diagnosis is very important for the treatment of brain damage.This review is to elucidate the relationship between imaging changes(CT,MRI and ultrasonography) and prognostic assessment,thus provide reference for clinicians,the objective is focus on imaging changes of HIE and intracranial haemorrhages,which are 3 of the most frequently reported cases.

Integrin is a kind of crucial adhesive molecule on the membrane,and it exerts its function through activating its intra-cellular effectors.One of the most important effectors is integrin-linked kinase (ILK),which is a serine/threonine protein kinase located in focal adhesions and has a key position in transducing many of the biochemical signals.It plays an important roles in regulating fundamental processes such as cell-matrix interactions,growth,proliferation,survival,differentiation,migration,invasion and angiogenesis under both physilogical and pathological processes.

OBJECTIVE To develop a method to construct three dimensional finite element model of nasomaxillary complex,in order to provide the foundation of studying nasomaxillary biomechanics characteristics.METHODS DICOM data from HelixCT were analyzed and reconstructed in software Mimics,then complete constructing three dimensional finite element model of nasomaxillary complex in Ansys.RESULTS The nasomaxillary complex model was similar to original case in structure,which is a high finite element model.CONCLUSION Computer assistant in constructing finite element model from DICOM data is a convenient,exact and efficient method .The nasomaxillay complex finite element model is exact and reproductable,and can provide the foundation of studying biomechanics mechanism of nasomaxillary fracture in otolaryngology.

OBJECTIVE:To investigate the anti-apoptosis effect and the mechanism of GM 1 ganglioside on nerve cell af?ter the spinal cord injury.METHODS:The rats with compressed injury at the T8.9level of spinal cord were employed as the model.Then the rats were divided into two groups at random,one for the control and one for GMI treatment.The rats in each group were administrated normal saline solution(20?l)and GM 1 (30?g,20?l)intrathecally10minutes after injury,respective?ly.The apoptosis of nerve cells in the injured spinal cord were examined by TUNEL and flow cytometry with nerve cells labeled with Annexin V/PI;and Caspase3activity was measured by fluorometric immunosorbent enzyme assay.RESULTS:The apoptotic cells appeared in both groups at4h and reached their peak on3rd day after the injury.The proportion of apoptotic cells and the intracelluar Caspase3activity in the GM 1 treated group were significant lower than those in the control group(P

The electrical pain perception were determined in healthy Chinese adults and factors were assessed. The anxiety-depression state of one hundred healthy Chinese were evaluated and the general information was recorded. Then electrical pain perception (perception threshold, pain threshold, pain tolerance) were assessed by neuromuscular monitor at the same time. Further study of influence factors such as gender, age, body mass index (BMI), left and right sides, degree of anxiety and depression, etc. were carried out on the pain perception. The results showed that 95% of the reference range of electrical perception threshold, pain threshold, pain tolerance were 4-7mA, 7 26mA and 15-60mA, respectively. The bilateral pain perception in males were significantly higher than that in females. There was no difference of effect between left and right sides. Age was positive related to pain perception, degree of depression was negative related to pain perception. BMI, degree of anxiety did not show association with pain perception. Therefore, gender, age and degree of depression were significantly related to electrical pain perception in healthy Chinese adults.
Adult ; Anxiety ; Body Mass Index ; Depression ; physiopathology ; Electric Stimulation ; Female ; Humans ; Male ; Pain Perception ; Pain Threshold ; Reference Values

BACKGROUND: The association between the ingredients of perfusate and its protection on corneal endothelium is always the hot issue in ophthalmology and pharmacology.OBJECTIVE: To observe the influence of different perfusates on the structure and function of rabbit corneal endothelium during phacoemulsification.DESIGN: A randomized grouping designed and controlled animal trial.SETTINGS: Laboratory of experimental animal center of Jinzhou Medical College and the laboratory of experimental animal operation of an urban hospital.MATERIALS: The experiments were carried out in the laboratory of experimental animal center of Jinzhou Medical College and the laboratory of experimental animal operation of Jinzhou Yadong Ophthalmology Hospital from September 2004 to March 2005. Sixteen pure Japanese big-ear rabbits of 3.5 months old, clean degree, were randomly divided into four groups with 4 rabbits in each group: normal control group, saline group, shike group and balanced salt solution group. Shike was produced by Shenyang Qixing Pharmaceutical Co.,Ltd.; Balanced salt solution by Alcon Company (USA).METHODS: The rabbits were intraperitoneally anesthetized with 100 ml/L chloral hydrate (3 mL/kg), 4% oxybuprocaine hydrochloride eye drops were used for surface anesthesia of eyes, and both eyes were operated. Alcon phacoemulsification apparatus (USA) and routine microsurgical instruments were used. A 3.5-mm incision was made on sclerotic tunnel at 2 mm posterior to superior limbus of sclera, punctured into the anterior chamber, then 0.25 mL Viscoat (Alcon) was infused. Curvilinear capsulorhexis was performed with the diameter of about 5 mm. The phacoemulsification head was placed in the center to suck out the crystal nucleus and cortex, and the incision was closed after the operation. The morphology of the corneal endothelium was quantitatively determined using contact specular microscope preoperativley and 6 hours postoperatively, including the density and area of corneal endothelium,percent of hexagonal cells and the coefficient of variation. At 6 hours postoperatively, trypan blue-alizarin red active staining was performed, and the changes of slight structures of corneal endothelium were observed under light microscope.MATN OUTCOME MEASURES: ① Quantitative analysis of the forms morphology of corneal endothelium (density and area of corneal endothelium, percent of hexagonal cells and the coefficient of variation); ② Characters of forms and structures of corneal endothelium.RESULTS: All the 16 rabbits were involved in the analysis of results. ① There were no significant differences in the morphological indexes among the four groups preoperatively. ② At 6 hours postoperatively, density and areas of endothelial cells, percent of hexagonal cells and the coefficient of variation were significantly lower in the saline group,shike group and balanced salt solution group than in the normal control group (P ＜ 0.05), and no significant difference was observed between the shike group and balanced salt solution group (P ＞ 0.05). ③ The structural changes of corneal endothelium in the shike group and in balanced salt solution group were alleviated more significantly than those in the saline control group, no necrosis was observed.CONCLUSTON: In phacoemulsification, the damage of perfusate to corneal endothelium is a chemical one. Under the same surgerical conditions, domestic perfusate of shike is as effective as balanced salt solution in protecting endothelial cells.

Thermostable L-arabinose isomerase (L-AI) is the most potential enzyme for the biological production of D-tagatose from D-galactose, a novel functional factor. Gene araA encoding the L-arabinose isomerase from Bacillus stearothermophilis IAM 11001 was cloned and expressed in Escherichia coli. The araA gene of 1491 bp has 95% identity with L-AI from Thermus sp. IM6501. The GenBank accession number for the nucleotide sequence of this araA gene determined in this work is EU394214.The bacterium was induced by IPTG and analyzed by SDS-PAGE, approximately 59 kDa exogenous protein was observed on the SDS-PAGE. The recombinant L-AI was purified to electrophoretical homogeneity with affinity chromatography, and the activity of recombinant L-AI was also studied. The bioconversion rate of D-galactose to D-tagatose reached 39.4% after 24h whole cell reaction.