Objective To study the expressions of S100B and glial fibrillary acidic protein(GFAP) in the central nervous system of Sprague-Dawley rats during postnatal development. Methods Twenty-four male SD rats were divided equally into four groups according to different postnatal times:7-day group,14-day group,21-day group and adult group.Immunohistochemistry was used to investigate the expressions of S100B protein and GFAP in the brain and the spinal cord. Results The amount and density of S100B positive astrocytes decreased significantly in frontal cortex,hippocampus,striatum,substantia nigra and spinal cord during postnatal development.It seemed that the second through the third week after birth was a critical period for these changes.The amount and density of GFAP positive AST increased gradually in the brain,but it was the opposite in the spinal cord.Double-labeled immunofluorescence of S100B and GFAP in hippocampus CA1 area showed that S100B positive stains were evenly distributed in the pyramidal,polymorphic and molecular layers from the seventh till the twenty first day after birth,but apparently decreased in each layer especially in the molecular layer in adult hippocampus while the immunoreactivities of GFAP increased.The proportion of double labeled cells also increased with the aging and more of them were found in the pyramidal and polymorphic layers.Conclusion Different patterns of the expressions of S100B and GFAP exist during postnatal development.It may reflect the different roles of these proteins on the glial cell development.And it may also indicate that the expressions of S100B and GFAP are regulated by different mechanisms during the course of development,which reflects the differentiation of subpopulations of astrocytes during ontogenesis.

Cochlear Implantation ; methods ; Cochlear Implants ; Models, Anatomic

Objective: To acquire recombinant nematode anticoagulant peptide ( NAP) with high anticoagulant activity. Methods: Pichia pastoris GS115 strain was transformed with recombinant yeast expression vector pPICS. 5K-rNAP. Expression of rNAP was induced with methanol after the identification of positive strains. NAP expressed in the collected yeast culture supernatant was confirmed with SDS-PAGE and Western blot. The biological activity of the products was validated with PT (prothrombin time) , INR (international normalized ratio) and APTT (activated partial thromboplastin time) , respectively. Results: The yeast strains expressing NAP were identified. The rNAP was secreted into culture supernatant with a molecular weight of about 10 kDa due to glycosylation, which is a little bigger than that predicted (8.7kDa). The anticoagulant efficiency of rNAP was confirmed with the in vitro assays. Conclusion: The recombinant nematode anticoagulant peptide with high biological activity was successfully expressed in Pichia pastoris and can be used in the future development of novel anticoagulant agent.

Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P ＜0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P＞0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.

Objective To explore the value of color doppler ultrasonography by bed side in the early diagnosis of HIE in full term neonates.Methods The changes of cerebral parenchymal and cerebral arterial blood stream parameter on 35 cases of neonates clinically diagnosed HIE of mild and moderate degree and 40 cases of normal newborns on the 24,48 and 72 hours after birth were observed by color doppler ultrasonography by bed side.Results 1.The cerebral parenchyma was even echo in normal newborns,but it was maldistributed and reinforced in mild asphyxia neonates and it was more serious in moderate degree.The echo of cerebral parenchyma in mild degree was near normal in 48 hours after birth,while the echo of cerebral parenchyma in moderate degree was still maldistributed and reinforced in 48 and 72 hours after birth.2.There was obvious changes in the cerebral arterial blood stream parameter and hemodynamics of the asphyxia newborns compared with normals.The systolic peak velocity(Vs)and end diastolic velocity(Vd)of the cerebral arteries in mild and moderate degree were obviously lower than that of control group in 24,48 hours after birth(Pa0.05).3.Resistance index(RI)of the cerebral arteries in mild and moderate degree were higher than that of control group in 24,48 hours after birth(Pa0.05).Conclusion Color doppler ultrasonography by bed side is a convenient,noninvasive method for diagnosing HIE.

Aged ; Genital Neoplasms, Male ; pathology ; Humans ; Male ; Neurilemmoma ; pathology ; Scrotum

To explore the effect of tripterygium glycosides on the level of peripheral blood cell factors of Graves ophthalmopathy (GO). In the study, 64 patients of GO in moderate-severe acute stage were selected, and randomly divided into the treatment group (32 cases) and the control group (32 cases). Both of the two groups were provided with basic treatment. The control group was added with prednisone(0. 75 mg kg-1 d-1 ), which gradually reduced (by 5-10 mg week-1 )to the minimum dose of 5 mg d-1. The treatment group was treated with 20 mg tripterygium glycosides, three times a day. One therapy course is three months. The levels of peripheral blood cells(TNF-alpha , IL-2, IL-10, IFN-gamma)of the two groups before and after the treatment and the clinical efficacy were observed. The study indicated that, before the treatment, TNF-alpha, IL-2, IFN-gamma in both groups were significantly higher than that in the health group, but with IL-10 notably lower than the healthy group. After the treatment, TNF-a, IL-2, IFN-gamma in the treatment group significantly decreased, but with IL-10 significantly increasing (P <0. 01). After the treatment, the two groups showed significant difference (P <0. 01). The total clinical efficacy in the treatment group was 88. 10% , and that in the control group was 57. 14% (P <0. 01). After the treatment, the two groups showed significant changes in the exophthalmos degree (P < 0. 01). The results showed that the level of peripheral blood cells (TNF-alpha,IL-2, IL-10, IFN-gamma)of GO patients was positively correlated with the severity of ocular disease. The combined therapy of tripterygium glycosides and methimazole show such advantages as low side effect and high clin-
Adult ; Cytokines ; blood ; Female ; Glycosides ; pharmacology ; therapeutic use ; Graves Ophthalmopathy ; blood ; drug therapy ; Humans ; Male ; Tripterygium ; chemistry

OBJECTIVETo establish the steriod-induced osteoporosis model with the type of Kidney-Yin deficiency.

METHODSTotally 45 female Kunming mice were randomly divided into normal group,model group and Liuwei Dihuang pills(Chinese character: see text)group. The model was established by intramuscular injecting of Dexamethasone. Liuwei Dihuang pills (Chinese character: see text) group was administered orally with Liuwei Dihuang pills (Chinese character: see text). The signs and symptoms of mice were observed dynamically. All the animals were sacrificed at the end of the 6th weeks. The level of ACTH, cAMP, cGMP, TSH and E2 in serum were detected to evaluate deficiency of Kidney-Yin. Morphological changes and bone density were observed to evaluate osteoporosis.

RESULTS(1) Compared with control group, mice in model group appeared obvious Kidney-Yin deficiency symptoms, including hair dry, restlessness, excitability, hard stool, and yellow. (2) Compared with control group,the weight of mice in model group gained slower (P<0.01); the index of adrenal gland,liver and spleen decreased (P<0.01, P<0.01 ,P<0.01); the level of ACTH and TSH increased (P<0.01 ,P<0.01), the level of E2 decreased (P<0.01) and the ratio of cAMP/cGMP increased (P< 0.05). (3)Compared with control group,the bone density of lumbar vertebra and femur in model group were significantly decreased (P<0.01, P<0.05); HE staining revealed osteoporosis in model group mice. (4)However, the Liuwei Dihuang pills (Chinese character: see text) group can partly antagonize the inhibition of the HPA axis, alter the disordered sex hormone and the ratio of cAMP/cGMP, and reverse the osteoporosis partly.

CONCLUSIONthe model of osteoporosis with type of Kidney-Yin deficiency could be established by Dexamethasone intramuscular injection. With less interference, it wight be a stable and reliable modeling method for integration of disease and syndrome in TCM.

Animals ; Bone Density ; Dexamethasone ; toxicity ; Disease Models, Animal ; Female ; Kidney Diseases ; etiology ; Medicine, Chinese Traditional ; Mice ; Osteoporosis ; chemically induced ; Yin Deficiency ; complications

Microscopy, Electron, Transmission ; methods ; Negative Staining ; methods ; Viruses ; ultrastructure

Chronic osteomyelitis is one of the most common disorder in clinic. In recent years due to diabetes, peripheral vascular disease and trauma induced disease increased, the prevalence rate increased. With the development of magnetic resonance imaging and CT imaging technology, it greatly improved the accuracy of clinical diagnosis of chronic osteomyclitis and ability to describe the infection characteristics, and provide a reliable basis for clinical treatment. The current research on chronic osteomyelitis mainly concentrated on the aspects of imaging applications and ways of using antibiotic optimization control inflammation, defect restoration and reconstruction of blood supply and treatment. But the best time to the antibiotic therapy and the use of program is still uncertain, for after debridement, bone grafting time and defect repair function of fast recovery still need further research.
Anti-Bacterial Agents ; therapeutic use ; Chronic Disease ; Humans ; Osteomyelitis ; diagnosis ; therapy