OBJECTIVE:To investigate the improving effect of phenylethanoid glycosides (PhGCs) from Tibetan medicine Phlomis younghusbandii on rats with acute high-altitude cerebral edema. METHODS:60 Wistar rats were randomly divided into a normoxia control group (isometric sterile water for injection),a hypoxia model group (isometric sterile water for injection),a dexamethasone group(4 mg/kg),and three groups of PhGCs at high(400 mg/kg),middle(200 mg/kg)and low(50 mg/kg)dos-es,with 10 rats in each group. The rats were given drugs,ig,6 d before the establishment of models. On the 4th day of administra-tion,ig,the rats in all groups except the normoxia blank group were placed in a simulated 8 000 m altitude plateau environment for 72 h hypoxic exposure to establish the rat models of high-altitude cerebral edema. Following HE stain,the pathological changes in rats’brain tissues were observed under the light microscope. Dry-wet proportion method was used to determine the water con-tents in rats’brain. The content of MDA and the activities of SOD and GSH in rats’brain tissues were detected. Enzyme-linked im-munosorbent assay was adopted to determine the contents of IL-1β and TNF-α in rats’serum and brain tissues. RESULTS:Com-pared to the rats in the normoxia control group,those in the hypoxia model group showed obvious brain edema,and thickened lacu-nas around cells and vessels and inflammatory cell infiltration, higher water contents and MDA and weaker activities of SOD and GSH in brain,and higher contents of IL-1β and TNF-α in serum and brain tissues. There were statistically significances (P<0.01 or P<0.05). Compared to the rats in the hypoxia model group,those in the groups of PhGCs at high,middleand low dosages demonstrated less inflammatory cell infiltration and lower water contents in brain tissues,in which the groups of PhGCs at high and middle dosages demonstrated lower content of MDA and stronger activities of SOD and GSH in brain tissues, and lower contents of IL-1β and TNF-α in serum and brain tissues. There were statistically significances (P<0.01 or P<0.05). CONCLUSIONS:PhGCs can obviously alleviate the acute cerebral injury in rats which is caused by acute hypoxia and has im-provement effect to some degree on the rats with acute high-altitude cerebral edema.

OBJECTIVETo isolate and identify the chemical constituents from Verbena officinalis.

METHODThe compounds were isolated by means of chromatography and the structures were determined on the basis of physical and spectral analysis.

RESULTFour compounds were isolated and identified as apigenin (I), 4'-hydroxywogonin (II), verbenalin (III) and hastatoside (IV).

CONCLUSIONCompounds I and II were obtained from the genus for the first time.

Apigenin ; chemistry ; isolation & purification ; Flavanones ; chemistry ; isolation & purification ; Iridoid Glycosides ; Iridoids ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Verbena ; chemistry

OBJECTIVETo study the effect of hyperbaric oxygenation (HBO) on the differentiation of the implanted human neural stem cells (hNSCs) into neurons in neonatal rats following hypoxic-ischemic brain damage (HIBD).

METHODSHIBD model was prepared by ligation of the left common carotid artery, followed by 8% hypoxia exposure in 7-day-old Sprague-Dawley rat pups. Three days later, the rats received implantation of hNSCs into the left cerebral ventricles. Then the survived rats were randomly divided into two groups: transplantation alone and transplantation+HBO (n=8 each). HBO treatment was administered (1.8 ATA, 1 hr once daily for 10 days) in the transplantation+HBO group 1 hr after hNSCs transplantation. Brains were removed 10 days after transplantation. Frozen coronal sections were prepared for immunofluorescence analysis to detect the neural differentiation of the transplanted cells in the cerebral cortex and hippocampus.

RESULTSDifferentiated neurons of implanted cells distributed mainly in the cortex and the hippocampus of the injured side. There was no difference in the number of neurons in the cortex between the two groups, while the number of neurons in the hippocampus significantly increased in the transplantation+HBO group compared with that in the transplantation alone group (231.4+15.1 vs 162.6+5.6; P<0.05).

CONCLUSIONSHBO treatment may promote the differentiation of implanted hNSCs into neurons in the hippocampus of neonatal rats following HIBD.

Animals ; Animals, Newborn ; Cell Differentiation ; Female ; Humans ; Hyperbaric Oxygenation ; Hypoxia-Ischemia, Brain ; therapy ; Male ; Neurons ; cytology ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation

OBJECTIVETo study the possible mechanism of aqueous extract of detoxified cottonseeds (CTN-W).

METHOD AND RESULTCTN-W 0.01, 0.03, 0.10, 0.30 mg.mL-1 was incubated directly with the synaptic membrane extracted from the cerebral cortex in rats, and adenylyl cyclase (AC) activity was detected by using radio-immunoassay.

RESULTShowed that CTN-W could activate AC in a dose-dependend manner. After incubation with PC12 cells in the presence of corticosterone 2 x 10(-4)mol.L-1 for 48 h, CTN-W 0.08, 0.4, 2 mg.mL-1 protected PC12 cells from the lesion induced by corticosterone.

CONCLUSIONAntidepressant and anxiolytic effects of CTN-W are related with the activation of AC-cAMP pathway in signal transduction system, thus protecting neurons from the lesion. These two aspects maybe partly form the mechanism of CTN-W's action.

Adenylyl Cyclases ; metabolism ; Animals ; Anti-Anxiety Agents ; pharmacology ; Antidepressive Agents ; pharmacology ; Corticosterone ; antagonists & inhibitors ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gossypium ; chemistry ; Neuroprotective Agents ; pharmacology ; PC12 Cells ; drug effects ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Seeds ; chemistry ; Synaptic Membranes ; enzymology

OBJECTIVETo analyze the influence of optimal codon usage on the expression levels and immunogenicity of DNA vaccines, encoding the human papillomavirus type 6b (HPV 6b) E7 gene.

METHODSThe full length E7 gene of HPV 6b was modified to substitute human preferred codon for rarely used codon, and three mutations were introduced into the pRB binding site of HPV 6b E7 to eliminate its transformation potential. The codon optimized and mutated E7 gene (hu-mE7) were cloned into the Kpn I and EcoR I site of the pcDNA3 mammalian expression vector, the in vitro expression of the hu-mE7 gene and the immunogenicity of hu-mE7 DNA vaccine were compared with the wt-E7gene.

RESULTSThe in vitro expression of pcDNA3-hu-mE7 was much higher than the classical wt-E7 plasmid in monkey COS-1 cell line. Mice immunized intramuscularly with the pcDNA3-hu-mE7 showed that the codon modified E7 gene induced a stronger IFN-gamma ratios than the wt-E7 gene.

CONCLUSIONSThese results suggest that the optimized codon usage contributes to the enhancement of gene expression and immunogenicity of HPV 6b E7 gene.

Animals ; Cell Line ; Codon ; genetics ; Female ; Gene Expression ; Genes, Viral ; genetics ; Genetic Vectors ; Mice ; Papillomaviridae ; genetics ; Papillomavirus Vaccines ; Transfection ; Vaccines, DNA ; immunology ; Viral Proteins ; biosynthesis ; genetics ; Viral Vaccines ; immunology

Objective To quantitative the changing information of estrogen receptor βgene which was in tissue and organ of sex gland during oestrus and dioestrus of Beagles, and to show the different expression situation of hypothalamus-pituitary-gonad axis during oestrus and dioestrus, and providing the basic of theory to research deeply the mechanism of heat of Beagles. Methods As the key gene in regulation reproduction, ERβgene is located in hypothalamus-pituitary-gonad axis, so using Beagles which was in oestrus and dioestrus, and extract the RNA from hypothalamus、pituitary、ovary and uterus respectively,after reverse transcription we detected the expression of ERβgene by real-time quantitative PCR.Results The expression of ERβgene mRNA from ovary、uterus、pituitary、hypothalamus of Beagles which was in dioestrus was 0.35 times, 0.17 times, 0.44 times and 0.43 times than the expression of ERβgene mRNA from ovary, uterus, pituitary, hypothalamus of Beagles which was in oestrus.Conclusion The expression of ERβgene was up-regulation in hypothalamus-pituitary-ovary axis of Beagles which was in oestrus.

OBJECTIVETo establish a RP-HPLC method for determination of glycosides in Traditional Chinese Medicine Liu-wei Di-huang.

METHODThe samples were analyzed on an ODS column at 30 degrees C, with mobile phase of methanol/water (33:67) at flow rate 1.0 mL.min and detection at wavelength of 236 nm.

RESULTThree major components reached base-line separation and were identified to be mononiside, loganin, paeoniflorin. Respectively for the three components, linear correlations were found between peak areas and concentrations in the ranges of 7.4-60, 7.7-62 mg.L-1 and 8.5-68 mg.L-1, and the recoveries were 98.8%, 98.3%, 99.6%.

CONCLUSIONThe established method is proved to be suitable for simultaneous quantification of three major glycosidic components in Liuwei Dihuang decoction and can be used for evaluation of the quality of Liuwei Dihuang preparations.

Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Chromatography, High Pressure Liquid ; methods ; Cornus ; chemistry ; Dioscorea ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; analysis ; chemistry ; Glucosides ; analysis ; Iridoids ; analysis ; Monoterpenes ; Plants, Medicinal ; chemistry ; Quality Control ; Rehmannia ; chemistry

OBJECTIVETo construct human-SCID chimeric mice through implantation of mononuclear cells from human cord blood and study the immunoreaction of SCID-Hu IC mice immunized with rAd5HPV16L1-E7 vaccine.

METHODS(1) Experiment groups were injected with the suspension of mononuclear cells from human cord blood through a tail vein; the control ones were injected with non serum RPMI 1640 medium. Eight weeks after implantation, blood was collected and human serum IgG level in the mice were tested, and human CD45, CD3 and CD19 were determined. (2) SCID-Hu IC mice were divided into two groups: in group A the mice were immunized intraperitoneally with rAd5HPV16L1-E7 virus and in group B the mice were immunized through nasal drip with rAd5HPV16L1-E7 virus. At the end of fourth week, the serum specific IgG antibody to rAd5HPV16L1-E7 virus, IFN-gamma in culture medium of spleen lymphocyte and T-lymphocyte propagation were tested.

RESULTS(1) In the experiment groups, the number of mice positive for human IgG was 10/15, the average values of CD45, CD3 and CD19 were (9.39+/-4.21), (3.25+/-3.99) and (1.69+/-0.75), respectively. In the control ones, the human IgG, CD45, CD3 and CD19 were negative. (2) The results in the experiment groups showed that the IFN-gamma and T-lymphocyte stimulated by HPV16 protein were higher than those in the non-stimulated group (P less than 0.05).

CONCLUSION(1) The results indicated that the construction of human-SCID chimaera through the implantation of mononuclear cells from human cord blood into SCID mice was successful. They also indicated that the reconstructed SCID-Hu IC mice has the ability to produce immune response against rAd5HPV16L1-E7 recombinant virus.

Adenoviridae ; genetics ; Animals ; Antigens, CD19 ; blood ; CD3 Complex ; blood ; Disease Models, Animal ; Female ; Fetal Blood ; transplantation ; Immunoglobulin G ; blood ; Interferon-gamma ; metabolism ; Leukocyte Common Antigens ; blood ; Male ; Mice ; Mice, SCID ; Oncogene Proteins, Fusion ; genetics ; immunology ; Oncogene Proteins, Viral ; genetics ; immunology ; Papillomaviridae ; genetics ; Recombination, Genetic ; T-Lymphocytes ; cytology ; Viral Vaccines ; immunology

Coronary artery disease (CAD) is a complex human disease, involving multiple genes and their nonlinear interactions, which often act in a modular fashion. Genome-wide single nucleotide polymorphism (SNP) profiling provides an effective technique to unravel these underlying genetic interplays or their functional involvements for CAD. This study aimed to identify the susceptible pathways and modules for CAD based on SNP omics. First, the Wellcome Trust Case Control Consortium (WTCCC) SNP datasets of CAD and control samples were used to assess the joint effect of multiple genetic variants at the pathway level, using logistic kernel machine regression model. Then, an expanded genetic network was constructed by integrating statistical gene–gene interactions involved in these susceptible pathways with their protein–protein interaction (PPI) knowledge. Finally, risk functional modules were identified by decomposition of the network. Of 276 KEGG pathways analyzed, 6 pathways were found to have a significant effect on CAD. Other than glycerolipid metabolism, glycosaminoglycan biosynthesis, and cardiac muscle contraction pathways, three pathways related to other diseases were also revealed, including Alzheimer’s disease, non-alcoholic fatty liver disease, and Huntington’s disease. A genetic epistatic network of 95 genes was further constructed using the abovementioned integrative approach. Of 10 functional modules derived from the network, 6 have been annotated to phospholipase C activity and cell adhesion molecule binding, which also have known functional involvement in Alzheimer’s disease. These findings indicate an overlap of the underlying molecular mechanisms between CAD and Alzheimer’s disease, thus providing new insights into the molecular basis for CAD and its molecular relationships with other diseases.

OBJECTIVETo evaluate breast reconstruction with a combined skin flap of the deep inferior epigastric perforator (DIEP) and the transverse rectus abdominis musculocutaneous (TRAM).

METHODSThe DIEP and TRAM united flap was elevated with the vessel pedicle of the deep inferior epigastric perforator on the affected side and the rectus abdominis muscle pedicle on the intact side. The reconstructive breast was shaped after the deep inferior epigastric vessels were anastomosed to the internal mammary vessels or the thoracodorsal vessels ipsilaterally.

RESULTSWe have used the DIEP and TRAM united flaps for breast reconstruction in 17 cases. All of the flaps survived, and the reconstructed breasts were well-shaped with the follow-up of 6-18 months.

CONCLUSIONSThe DIEP and TRAM united flap possesses of advantages such as rich blood supply, abundant tissue volume and easy shaping. It is especially applicable to the cases who have large chest defect and need large volume tissue.

Adult ; Epigastric Arteries ; transplantation ; Female ; Humans ; Mammaplasty ; methods ; Middle Aged ; Rectus Abdominis ; transplantation ; Skin Transplantation ; Surgical Flaps ; blood supply