The M and NP genes of H5N1 avian influenza virus (A/chicken/Hubei/489/2004) were amplified by RT-PCR from viral RNA,and cloned into pMD 18-T vector respectively.The expression plasmid containing the M gene (pHM6-m) or the NP gene (pHM6-np) was then constructed by inserting the M or NP gene into the pHM6 eukaryote expression vector; the constructed plasmid was then sequenced.32 BALB/c mice (6-week-old) were divided into four groups at random.Three groups of BALB/c mice were inoculated one time the intramuscular route with either 30 μg of plasmid pHM6-m,30 μg of plasmid pHM6-np or the mixture of plasmid pHM6-m (15 μg ) and pHM6-np(15 μg) respectively.A additional group of mice were injected with 100 μ1 PBS as controls.Two weeks later,all mice were challenged with homologous H5N1 avian influenza virus,and observed in the following 12 days.The survival rates of mice in the pHM6-m group,the pHM6-np group and mixed plasmids group were 62.5% ,25.0% and 50.0%,respectively.Results showed that effective protection could be provided by either pHM6-m or pHM6-np,but pHM6-m provided a better protective effect than pHM6-np.

Objective :To observe influence of Ginkgo ketone ester dropping pills (GKE) combined atorvastatin on vascular endothelial function ,serum levels of omentin-1 and homocysteine (Hcy) in hypertensive patients with ca-rotid arteriosclerosis (CAS).Methods : A total of 120 hypertensive patients with CAS treated in our hospital were selected ,randomly and equally divided into atorvastatin group and combined treatment group (received GKE com-bined atorvastatin ) ,both groups received corresponding therapy based on health education ,diet and exercise etc for 12 weeks.Blood pressure ,blood lipids ,carotid intima-media thickness (IMT ) ,plaque size ,levels of endothelin (ET)-1 ,nitric oxide (NO) ,serum omentin-1 and Hcy were compared between two groups before and after treat- ment .Results : Compared with before treatment ,there were significant reductions in blood pressure ,blood lipids (except significant rise in HDL-C) ,serum levels of ET-1 and Hcy ,IMT and plaque size ,and significant rise in ser-um levels of NO and omentin-1 in two groups after treatment , P＜0-01 all.Compared with atorvastatin group after treatment ,there were significant reductions in blood pressure ,blood lipids (except significant rise in HDL-C) ,ser-um levels of ET-1 [ (136-42 ± 15-57) μg／ml vs.(116-57 ± 13-28) μg／ml] and Hcy [ (15-46 ± 3-25 ) μmol／L vs. (10-58 ± 3-11) μmol／L] ,IMT [ (1-43 ± 0-21) mm vs.(1-26 ± 0-19) mm] and plaque size [ (22-53 ± 1-28) mm2 vs.(20-67 ± 1-14 ) mm2 ] ,and significant rise in serum levels of NO [ (102-43 ± 12-37) μmol／L vs.(129-84 ± 13-36) μmol／L] and omentin-1 [ (176-57 ± 45-20 ) μg／ml vs.(198-73 ± 45-38) μg／ml] in combined treatment group , P＜0-05 or ＜0-01- Conclusion : Ginkgo ketone ester dropping pills combined atorvastatin can significantly improve vascular endothelial function and arteriosclerosis while reducing blood pressure and blood lipids in hyperten-sive patients with carotid arteriosclerosis .

Objective To investigate the significance of the serum levels of interleukin-10 (IL-10),IL-13,IL-15 of patients with chronic hepatic failure and the correlation between those inter- leukin levels and nosocomial infections.Methods The serum levels of IL-10,IL-13,IL-15 of 58 patients with chronic hepatic failure were measured by double antibody sandwich enzyme-linked immu- nosorbent assay at the time of admission and 2 weeks after admission.Results The serum levels of IL-15 and the propotion of IL-15/IL-10 and IL-15/IL-13 in patients with chronic hepatic failure group at the time of admission were significantly higher than those in healthy control group[(358.16?290.91) ng/L vs (38.55?21.49) ng/L,12.93?14.26 vs 1.10?0.55,98.55?97.5.5 vs 9.70?5.03,respectively,all P=0.000].Those in death group were significantly higher than those in improving group[(479.93v205.52) ng/L vs (244.51?236.29) ng/L,17.65?17.78 vs 8.53?7.98,130.69?115.50 vs 68.55?65.99,respectively,all P

Objective To study the expression and regulation of polycystic ovary syndrome ( PCOS) susceptibility gene Hmga2 in endometrial receptivity and decidualization. Methods Experiments including early pregnancy, delayed implantation and activation, artificial decidualization and ovarian steroid hormone treatment were performed, and Q-PCR and Western blot analyses were applied to explain the role of Hmga2 in endometrial receptivity. Results The Hmga2 ex-pression increased gradually with pregnancy. Its expression at the implantation site was significantly higher than that at the inter-implantation site,in the embryo activation group than in delayed implantation group, and in the artificial decidualiza-tion than the non-decidualization. The expression of Hmga2 was positively correlated with the expression of estrogen and progesterone, and was positively regulated by estrogen and progesterone in vivo. Conclusions Our findings indicate that the expression of Hmga2 is closely related to the embryo implantation process in early pregnancy in mice, is involved in the process of endometrial decidualization, and is influenced by the activated blastocyst and steroid hormones.

OBJECTIVEIn order to investigate the characterization of mother to children transmission, the sequences of HBV were analyzed to offer the information about the effect of interrupt.

METHODSThe sera of 75 mother with positive HBsAg are collected from 2003, and the ELISA was performed to determine the HBV infection of the child. The Large S sequence of HBV including preS and S gene are amplified and sequenced. The genotype was determined with the standard genotype sequence. The mutation ratios of group successfully interrupted and failed compared.

RESULTSThe sera of 4 pairs mother-children were HBsAg positive, including one twins. The virus genes are successful amplified. Four of HBV genotype is B and one is C. Gene of twins has mutation of T143M. 43 HBV of successfully interrupted group were sequenced. There are 37 of genotype B and 6 of genotype C. Three have the mutation in "a" dominant, and the percentage is 7%.

CONCLUSIONMost failed interrupted child have the same sequence with their mother, and the ratio is higher than the mother of successful group, however there have no statistical significance.

Adult ; Female ; Hepatitis Antibodies ; blood ; Hepatitis B ; immunology ; prevention & control ; transmission ; virology ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Hepatitis B virus ; genetics ; immunology ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; immunology ; Infectious Disease Transmission, Vertical ; prevention & control ; Male ; Mutation ; Vaccination

Adult ; China ; epidemiology ; GB virus C ; classification ; Hepatitis, Viral, Human ; epidemiology ; virology ; Homosexuality, Male ; Humans ; Male

OBJECTIVETo investigate the biological characters of human skin fibroblasts in fibroblast populated collagen lattice (FPCL).

METHODSThe human fibroblasts were cultured in 3D and the collagen of the rat tail was also prepared. They were examined with the comprising cell cycle and apoptosis, mRNA expression of TGF beta1, and fibronectin, and cell morphology.

RESULTSThe flow cytometry showed that the G0/G1, stage cells were 79% +/- 3%, 87% +/- 2% after the 7 days and 14 days separately, and there were not apoptosis peak observed. RT-PCR analysis revealed that the mRNA expression of TGF beta1, and fibronectin had no difference between human skin fibroblasts cultured in 3D and 2D. Electron microscope showed the cells were plenty of chromatin and organelles.

CONCLUSIONSThe proliferation of the human skin fibroblasts in FPCL is slow, but its biological viability is better.

Animals ; Cell Culture Techniques ; Cell Division ; Cells, Cultured ; Collagen ; Extracellular Matrix ; Fibroblasts ; cytology ; Humans ; Rats ; Skin ; cytology ; Tissue Engineering ; methods

OBJECTIVETo investigate whether mammalian target of rapamycin (mTOR) kinase was abnormally activated in maldeveloped balloon cells and dysmorphic neurons of focal cortical dysplasia (FCD) with refractory epilepsy.

METHODSA total of 12 archival cases of FCD typeIIwith medically intractable epilepsy treated between 2008 and 2010 were retrieved. Perilesional brain tissue was used as control specimens (n = 8). The expression of phosphorylated p-AKT (Ser473), p-mTOR (Ser2448) and p-P70S6K (Thr389) was investigated by imunocytochemistry.

RESULTSThe expression of p-AKT (Ser473), p-mTOR (Ser2448) and p-P70S6K (Thr389) was found in meldeveloped balloon cells and dysmorphic neurons of FCD. A weak stain in a small amount of pyramid neurons was also found in the control group.

CONCLUSIONAbnormal activation of mTOR in maldeveloped balloon cells and dysmorphic neurons of FCD may be a key molecular mechanism underlying the histological changes and repeated seizures.

Adolescent ; Adult ; Brain Diseases ; metabolism ; pathology ; Child, Preschool ; Epilepsy ; metabolism ; pathology ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Immunohistochemistry ; Male ; Malformations of Cortical Development ; metabolism ; pathology ; Malformations of Cortical Development, Group I ; Nestin ; metabolism ; Neurons ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; TOR Serine-Threonine Kinases ; metabolism ; Young Adult

OBJECTIVETo explore the effective and safety of carotid endarterectomy (CEA) in the treatment of severe stenosis of internal carotid artery.

METHODSThe clinical data of 95 patients with internal carotid artery stenosis and treated by CEA in our hospital from October 1998 to October 2006 were retrospectively analyzed. Transient ischemic attack (TIA) was found in 65 patients and cerebral infarction in 21 patients before operations. Diagnoses were confirmed by selective angiography, and > 70% stenosis of internal carotid artery was found in all patients and > 95% stenosis was found in 50 patients. Stenosis or occlusion of the contralateral carotid artery was also found in 35 patients. Coronary artery stenosis was confirmed in 43 patients with preoperative coronary angiography. CEA and coronary artery bypass grafting (CABG) were simultaneously performed in 18 patients.

RESULTSAfter the operation, the clinical symptoms significantly improved in all patients. One case experienced hypoglossal nerve injury. One patient died of heart attack during follow-up.

CONCLUSIONCEA is an effective and safe treatment for severe stenosis of internal carotid artery.

Aged ; Aged, 80 and over ; Carotid Artery, Internal ; surgery ; Carotid Stenosis ; surgery ; Endarterectomy, Carotid ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies

Objective To observe Th-1 drift induced in vitro by high immunogenic glioblastoma multiforme (GBM) U251 cell vaccine with high expression of membrane-enriched heat shock protein 70 (Hsp70) and major histocompatibility complex class Ⅰ (MHC-Ⅰ) molecules. Methods The high expression of MHC-Ⅰ and Hsp70 in U251 cells were induced by 500 U/ml IFN-γ for 48 h, heat shock at 43 ℃ for 2 h, or their combination. The cells were then inactivated by the mitomycin C (MMC) to prepare the cell vaccine. Peripheral blood mononuclear cells (PBMCs) from healthy donators were incubated with GBM U251 cell vaccines as the effector cells. Flow cytometry was applied to analyze the changes of CD4<'+> and CD8<'+>T lymphocytes in the PBMCs. The secretion of IFN-γ and IL-2 of the effector cells, after assaulting the target cells, was evaluated by ELISA. Results The percentages of CD4<'+> and CD8<'+> T lymphocytes of the PBMCs incubated with the U251 cell vaccine increased significantly as compared to that stimulated by the membrane-enriched MHC class Ⅰ or Hsp 70 molecule U251 cell vaccines (P<0.05), and so was the secretion of IFN-γ and IL-2 (P<0.05). Conclusions Th-1 drift stimulated by GBM U251 cell vaccine with high immunogenicity, high expression of Hsp 70 and membrane-enriched MHC class Ⅰ molecules plays an important role in antitumor mechanism in vitro.