Objective:To identify the expression of egfp-hri fusion on NIH3T3 cells which is carried by recombinant retrovirus. Methods:The egfp-hri fusion gene was integrated intoNIH3T3 cells by the infection of recombinant viral supernatant from the G418 resistent PA317 cells transfected stably with the plasmid of pLNCX-EGFP-C1-hri. The expression effect of egfp-hri fusion was visualized under the fluorescent microscope, while the expression levels of egfp-hri fusion protein by Western blot. Results:The observation from fluorescent microscope showed the green fluorescent was obviously observed in the cytoplasm , western-blotting showed the expression level of egfp-hri fusion gene increased by 83%and 81%, as compared with those uninfected cells and in the cells infected with empty vector, respectively. Conclusion:The infected NIH3T3 cells can express egfp-hri fusion protein successfully.

Objective To explore mechanism of epigallocatechin-3-gallate (EGCG) on improvement of cognitive function and alleviation of hippocampal insulin resistance in APP/PS 1 transgenetic mice. Method 12 months old female APP/PS 1 mice were randomly divided into 3 groups:model group(Tg), EGCG low dose group (Tg/EGCG-L), high dose group(Tg/EGCG-H). C 57 BL/6 J mice were utilized as control. learning and memory ability in 4 group mice were detected by morris water maze test(MWM). The hippocampal TNF-α/JNK signal and IRS-1 pSer 312 expression were detected by Western blot and immunohistochemical staining. Results Compared with NT mice, Tg mice showed a marked prolongation of the escape latency and swimming distance in the MWM test(P<0.05);Abnormal activation of TNF-α/JNK signaling and increased IRS-1 pSer 312 expression in the hippocampus of Tg mice(P<0.05). EGCG-treated Tg mice showed significantly improvement of all these abnormal changes(P<0.05). Conclusion EGCG treatment is able to alleviate hippocampal insulin resistance and improve cognitive function in the APP/PS 1 mice. which may be partly attributed to the reduction of TNF-α/JNK signaling activity in this AD mouse model.

Coupled magnetic nanoparticles in the microcapsule structure, such as magnetic microcapsules, can be delivered in specific organism or tissues under magnetic field exposure. Thus, the microcapsules can achieve active targeting functions by manipulat-ing the magnetic field. Based on the magnetic microcapsules, the antitumor drugs can also be loaded to realize magnetic response, which gives microcapsules sustained and controlled release advantages. To date, the drug microcapsules carrying magnetic nanoparti-cles have become promising novel delivery carriers for the treatment of tumor diseases. This paper mainly reviews the method of prepa-ration of the magnetic nanoparticle-coupled microcapsules, including liposomes, polyelectrolyte microcapsules, and polymer micro-spheres. The basic research progress of these microcapsules as anticancer drug carriers for the tumor therapy was also reviewed.

Objective:To identify the expression of plasmid pLNCX-EGFP-C1-hri targeting the gene of Human ribonuclease inhibitor (hri) in PA317 cells which is capable of expression in mammalian cells.Methods: The vector of pLNCX-EGFP-C1-hri was transfected into PA317 cells by Lipofectamine 2000 and then the expression of recombinated plasmid was verified in living cells by observing the transcription level of egfp-hri fusion gene mRNA with RT-PCR method and the expression level of egfp-fusion hRI protein with western blotting method respectively.Results: Both RT-PCR and western blotting showed the egfp-hri fusion gene was obviously expression in PA317 cells.Conclusion: The plasmid of pLNCX-EGFP-C1-hri targeting hRI is successfully constructed and the protein of hRI can be expressed in PA317 cells correctly.

OBJECTIVETo explore the effect and mechanism of cinnabaris and realgar in promoting awake effect of endotoxin- induced brain injury rat applied with Angong Niuhuang Wan.

METHODNormal rats implanted cortical electrode in advance were divided into 6 groups: control, model, the Angong Niuhuang Wan (AGNH, 0.4, 0.2 g · kg(-1)), the Angong Niuhuang Wan without cinnabaris and realgar (QZX-AGNH, 0.32, 0.16 g · kg(-1)). Rats in the control and model groups were given distilled water. After three days of intragastric administration, the brain injury model was injected with endotoxin through tail vein. Then trace electro-corticogram (EcoG) 1-6 h after LPS injection, and compare the power and relative power of beta (β) and delta-waves (δ) at 6 h of these groups. The content of acetylcholine (Ach) and the affinity of M-receptor (M-R) in cortex and brainstem were detected by alkaline hydroxylamine colorimetric method and radioactive ligand binding assay, respectively.

RESULTAGNH (0.4, 0.2 g · kg(-1)) could increase the power and relative power of β and AGNH (0.4 g · kg(-1)) showed better action on brain electrical activation. QZX-AGNH showed weak effect on it. AGNH (0.4 g · kg(-1)) could increase the affinity of M-R in cortex and the content of Ach in brainstem. The action of QZX-AGNH was not obvious.

CONCLUSIONIn endotoxin-induced brain injury rats, AGNH can raise the cholinergic system function of cortex, and strengthen the uplink of cortex activation of brainstem cholinergic system, improve the level of cortical activity and enhance the activation of EcoG to promote the body's awakening. QZX-AGNH show weak effect. Cinnabaris and realgar play an important role in promoting awake effect in endotoxin-induced brain injury applied with Angong Niuhuang Wan. The mechanism may be related to cortical and brainstem cholinergic system function.

Animals ; Brain Injuries ; chemically induced ; drug therapy ; physiopathology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; Endotoxins ; adverse effects ; Humans ; Male ; Rats ; Rats, Sprague-Dawley

Child ; Humans ; Motor Skills Disorders ; diagnosis ; therapy

Objective To explore the clinical and laboratory characteristics of langerhans cell histiocytosis (LCH) in children, so as to improve diagnosis level and decrease misdiagnosis rate.Methods Twenty-five cases of LCH from Jan.1996 to Feb.2006 were analyzed by retrospective study. The clinical data were collected and abstracted for information regarding clinical symptom, physical sign, laboratory examination, imaging,pathology,diagnosis and treatment.Results Some laboratory findings in hemogram, bone marrow examination and chest X-ray were non-specific.The X-ray characteristic of skeleton was osteolysis. Computerized tomography and magnetic resonance imaging were important in defining the extent of lesion in fundus cranii and sella.Seven cases were examined for anti-Epstein-Barr virus(EBV) IgM, 3 cases were positive;5 and 3 cases out of 10 cases showed humoral and cellular immunity abnormality,respectively. The misdiagnosis rate was 52%,1 case had been misdiagnosed for 7 years. Chemotherapy was effective in the future.Conclusions The clinical manifestations of LCH varies widely, leading to high rate of misdiagnosis.The etiology of LCH is unclear,and some of our patients show the evidence of EBV infection or immunity abnormality. Definitive diagnosis of LCH is based on pathology. Ultrastructure and immunophenotype should be done to improve diagnosis level.

This study taking gestodene (GEST) as a model, investigated the factors affecting reservoir-type intravaginal ring (IVR)'s drug release. This paper reported a gestodene intravaginal ring of reservoir design, comprising a gestodene silicone elastomer core encased in a non-medicated silicone sheath, separately manufactured by reaction injection moulding at 80 degrees C and heating vulcanization at 130 degrees C is reported. The test investigated the factors affecting drug release through a single variable method, taking the drug release rates of 21 days as standards. When changing the thickness of the controlling sheath outside, the ratio of the first day of drug release and mean daily release (MDR), named the relatively burst effect, is closing to 1 with the thickness of controlling sheath increasing, while the 1.25 mm sheath corresponding to 1.04 controlled the burst release effectively; a positive correlation (r = 0.992 2) existed between the average drug release (Q/t) and drug loading (A) within a certain range. The C6-165 controlling sheath with high solubility of GEST is easier to achieve controlled release of the drug; GEST crystalline power is more effective to implement controlled release of drugs among difficent states of the drug. A 1/4 fractional segment core gives a relatively burst effect of 1.76, while the 1/1 and 1/2 are 1.93 and 1.87 separately, at the same drug loading, concluding that use of a fractional segment core would allow development of a suitable GEST reservoir IVR. In summary, GEST reservoir-type IVR could be adjusted by the thickness of controlling sheath, the loading of drug, the material properties of controlling sheath, the dispersion state of drug, the additive composition and structure of intravaginal ring, to control the drug release behavior and achieve the desired drug release rate.
Administration, Intravaginal ; Contraceptive Agents, Female ; administration & dosage ; Contraceptive Devices, Female ; Delayed-Action Preparations ; Drug Delivery Systems ; methods ; Norpregnenes ; administration & dosage ; chemistry ; Silicone Elastomers ; chemistry ; Solubility

According to bias in codon choice of Pichia pastoris, The phytase phyA gene from Aspergillus niger N25 was mutated without changing its amino acid sequence. The expression plasmid pPIC9k-phyAm was constructed and transformed into GS115 strain. Positive clones,of which the chromosomes were integrated with phyA gene,were identified by the phenotype and PCR. SDS-PAGE analysis suggust that the size of enzyme protein of the expression product was about 70.15kDa.Southern blotting analysis to the yeast transformants showed that phyA gene was intergrated into the chromosome genome. The phytase activity of PP-NP m-4-4 with codons optimized reached 136 000U/ml in malt wort culture medium after being induced with 36h, which was the 2.8 times of the original strain PP-NPm-8.

Objective To alanyze the imaging features of brain in systemic NHL.Methods CT and MRI of brain in 8 cases of systemicNHL proved pathologically were performed.Imaging findings of brain were analysed and by comparison with that in 73 cases of brainmetastases with marked enhancement after contrast administration.Results 13 lesions in 8 cases were found by CT,the findings on contrast-enhanced CT included:①Ten lesions showed low density with obscure border;②Three nodular lesions showed high density(including 2 homogeneously and 1 ring-like enhanced nodules).All lesions showed neither ventricle enlargement nor occupying effects.Those enhanced showed no edema around the nodules.Of the 4 cases with MRI,low T_1 signal and high T_2 signal with no change after contrast administrationwere seen in 3 cases.One showed ring-like enhancement with no edema around.While 89% of the 73 brain metastatic cases showeddifferent extent of edema.Conclusion The imaging findings in some of the lesions in this study are different from that described before in literatures.In comparison with other metastases,the marked-enhanced nodules with no edema were found to be a specialty of secondary NHL in brain in our cases.