OBJECTIVETo observe the effect of Angong Niuhuang Pill (ANP) on Thl/Th2 in cerebral infarction patients and to explore the mechanism of phlegm-heat obstructing orifices.

METHODSRe- cruited were 30 cerebral infarction patients of phlegm-heat obstructing orifices syndrome (PHOOS) both in China and Indonesia. They were assigned to 4 groups according to the use of ANP, the Chinese treatment group, the Indonesia treatment group, the Chinese control group, and the Indonesia control group. Patients in the two control groups received conventional treatment, while those in the two treatment group additionally took ANP for 30 successive days. Their adverse reactions were observe, and levels of INF-γ and IL-4 were detected.

RESULTSThe INF-γ level and the INF-γ/IL-4 ratio significantly decreased, and the IL-4 level increased after treatment in the four groups with statistical difference (P < 0.05). Compared with the control group after treatment in the same country, the INF-γ level and the INF-γ/IL-4 ratio were lower, and the IL-4 level was higher in the two treatment groups (P < 0.05). Compared with the two Chinese groups, the INF-γ level and the INF-γ/IL-4 ratio were higher, and the IL-4 level was lower in the two Indonesian groups (P < 0.05). There was no statistical difference in the post-treatment indices between the two treatment groups.

CONCLUSIONSANP had moderating effect on Th1/Th2 in cerebral infarction pa- tients. Cerebral infarction patients of PHOOS might exist certain relation with Th1/Th2.

Cerebral Infarction ; drug therapy ; China ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hot Temperature ; Humans ; Indonesia ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism

Intersectin is an evolutionarily conserved multifunctional adaptor protein with multifunctional domains. These domains interact with components of the endocytic and exocytic pathways, such as the clathrin mediating synaptic vesicle recycling, the protein related to endocytosis via caveolae, the with-no-lysine kinases related to the regulation of renal outer medullar potassium, and the Cdc42 mediating exocytic pathway. Recently, the understanding of intersectin function in the pathogenesis of endocrine tumor and many neurodegenerative diseases such as Down syndrome, Alzheimer disease has been deepened. This article reviewed the structure and roles in endocytosis/exocytosis and diseases of intersectin.
Adaptor Proteins, Vesicular Transport ; physiology ; Endocytosis ; Exocytosis ; Humans ; Synaptic Vesicles ; physiology

1H NMR metabonomics approach was used to reveal the chemical difference of urine between patients with Xiao-Chaihu Tang syndrome (XCHTS) and healthy participants (HP). The partial least square method was used to establish a model to distinguish the patients with Xiao-Chaihu-Tang syndrome from the healthy controls. Thirty-four endogenous metabolites were identified in the 1H NMR spectrum, and orthogonal partial least squares discriminant analysis showed the urine of patients with Xiao-Chaihu Tang syndrome and healthy participants could be separated clearly. It is indicated that the metabolic profiling of patients with Xiao-Chaihu Tang syndrome was changed obviously. Fifteen metabolites were found by S-pot of OPLS-DA and VIP value. The contents of leucine, formic acid, glycine, hippuric acid and uracil increased in the urine of patients, while threonine, 2-hydroxyisobutyrate, acetamide, 2-oxoglutarate, citric acid, dimethylamine, malonic acid, betaine, trimethylamine oxide, phenylacetyl glycine, and uridine decreased. These metabolites involve the intestinal microbial balance, energy metabolism and amino acid metabolism pathways, which is related with the major symptom of Xiao-Chaihu Tang syndrome. The patients with Xiao-Chaihu Tang syndrome could be identified and predicted correctly using the established partial least squares model. This study could be served as the basis for the accurate diagnostic and reasonable administration of Xiao-Chaihu-Tang syndrome.
Humans ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; Metabolomics ; Proton Magnetic Resonance Spectroscopy ; Syndrome ; Urinalysis

OBJECTIVETo investigate the expression of Tenascin-C mRNA in keloids and hypertrophic Total RNA was isolated from normal adult skin. A cDNA fragment (base 5941-6481bp) of the scars.

METHODSfull-length human Tenascin-C cDNA was synthesized by polymerase chain reaction and subcloned in pGEM-T-easy. Dioxygen-labeled anti-sense and sense probes were synthesized by using a Sp6/T7 RNA synthesis kit in the present of Dig-UTP in vitro. The samples were taken from keloids in 10, hypertrophic scars in 10 and normal adult skin in 5. The hybridization was performed with 4% paraformaldehyde-fixed and wax-embedded sections to detect the Tenascin-C mRNA.

RESULTSThe Tenascin-C mRNA was negative in the normal adult epidermis and weakly located in the fibroblasts of the papillary dermis and the epidermal adnexa. In all of the 10 keloid specimens, the Tenascin-C mRNA was positive throughout the epidermis and widely distributed in the dermis included in the fibroblasts, endothelial cells and epidermal adnexa. In the specimens of the 3 hypertrophic scars,the Tenascin-C mRNA was also positive in the epidermis, but in the other 7 cases, it became negative. In the dermis of the hypertrophic scar,the Tenascin-C mRNA was weaker than that in the keloid, but stronger than that in the normal skin.

CONCLUSIONSThe expression of Tenascin-C mRNA is markedly enhanced in the keloids.

Cicatrix, Hypertrophic ; genetics ; metabolism ; pathology ; Female ; Humans ; Keloid ; genetics ; metabolism ; pathology ; Male ; RNA, Messenger ; metabolism ; Tenascin ; genetics ; metabolism

OBJECTIVETo investigate of antiatherogenic effect and possible mechanisms of piper longuminine.

METHODThe atherosclerotic model was established by the hypercholesterol feeding rabbits. Male Mew Zealand rabbits were randomly divided into five groups: normal group, model group, the high-dose (5 mg x kg(-1) x d(-1)) and low-dose (2.5 mg x kg(-1) x d(-1) group of piperlonguminine, and simvastatin group (5 mg x kg(-1) x d(-1)). All the rabbits were fed for 60 days. Blood samples were taken from the ear edge vein of rabbits in the day before the experiment, and in the days of 20, 40 and 60 days after the experiment, respectively. All the rabbits were fasted for at least twelve hours before the blood was taken. The blood serum were analyzed for total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C). The blood serum of the 60th day were also analyzed for superoxide dismutase (SOD), malondialdehyde (MDA) and nitric oxide (NO). At last, the pathological observation of aorta and heart samples were carried out.

RESULTCompared with those in model group, the TC, TG and LDL-C levels were reduced (P < 0.05) and the HDL-C was raised in the piperlonguminine group; also, the serum SOD and NO level was raised (P < 0.05), MDA level was reduced in the piperlonguminine group (P < 0.05). Area percentage of aorta plaque was reduced (P < 0.01) in the piperlonguminine group. The aorta and heart injury was abated and coronary artery angusty extent was markedly abatement (P < 0.01). The results of observation through transmission electron microscope (TEM) indicated that the fine structure of aortal pathological degree was markedly abated.

CONCLUSIONThe piperlonguminine could inhibit the atherogenesis formation and development, which might be due to regulating the lipid metabolism and enhancing the antioxidation.

Animals ; Atherosclerosis ; blood ; chemically induced ; drug therapy ; pathology ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Dioxolanes ; pharmacology ; Male ; Malondialdehyde ; blood ; Nitric Oxide ; blood ; Rabbits ; Superoxide Dismutase ; blood ; Triglycerides ; blood

ObjectiveTo investigate the long term clinical result of three-dimensional radiotherapy for esophageal carcinoma,discuss the effect of correlative factors to survival and local control.Methods From July 2003 to December 2008,792 patients with esophageal cancer were eligible.Patients were treated with three-dimensional radiotherapy (672 patients) or intensity-modulated (120 patients) radiotherapy.The radiotherapy was delivered in 1.8-2.0 Gy per fraction,5 fractions per week,total dose of 50-70 Gy,(median,60 Gy).142 patients were treated by concurrent radiochemotherapy,and the other 650 patients radiotherapy alone.The local control rate and survival rate were calculated by Kaplan-Meier method.Logrank method was used for univariate analyses.Cox regression model was used for multivariate analyses.ResultsThe follow-up rate was 95.8％.The number of patients with 5 years time followed-up was 133.The 1-year,3-year and 5-year local control rates were 76.6％,53.2％,48.6％,and the 1-year,3-year and 5-year overall survival rates were 70.1％,36.7％ and 28.0％,respectively.There were significant influence on the prognosis of T stage,N stage,TNM stage,tumor volume ( x2 =20.58-55.60,all P =0.000).The Cox multivariate model showed that N stage and tumor volume were independent prognostic factors (x2 =6.35,29.23,P =0.012,0.000).For the two groups of concurrent chemo-radiotherapy and radiotherapy alone,5-year local control rates were 57.0％ and 46.8％ ( x2 =7.34,P =0.007 ),the 5-year overall survival rate 32.8％ and 27.6％ ( x2 =3.42,P =0.064.ConclusionsThree-dimensional radiotherapy is effective for esophageal carcinoma.It might improve the local control rate and overall survival rate to some extent.T staging,N staging,TNM staging and tumor volume were important prognostic factors for long-term survival.The addition of concurrent radiochemotherapy could improve local control rates.

OBJECTIVETo explore the hematopoietic reconstitution potential of mesenchymal derived stem like cells.

METHODSWe transplanted bone marrow mesenchymal derived stem like cells into lethally irradiated BALB/c mice. Hematopoietic cells were derived from the non-adherent bone marrow cells 24 hours after initial culture while murine mesenchymal derived stem like cells from bone marrow of donor mice were cultured for 10 days before the transplantation.

RESULTSAll mice of group 1 and 3 died in 7-8 days post irradiation following transplantation, while all the mice from group 2 and 4 survived. The time course of hematopoietic reconstitution was then observed. The peripheral blood and bone marrow cell count recovered in the MSC + G-CSF transplanted group and the BM transplanted group after 3 weeks. Interestingly, CFU-GM number in the MSC + G-CSF transplanted group increased significantly after 2 weeks and even more than that in the BM transplanted group after 3 weeks while as CFU-GM colony dropped 2 weeks after in the BM transplanted group. Spleen colony (CFU-S) number and size of the MSC + G-CSF transplanted group was significantly greater than the BM transplanted group. Furthermore, PCR analysis was performed using peripheral blood cells to determine if any male-derived cells were present. No male-derived cells were found in any of the mice from group 1 and 3. Y-chromosome-specific src gene was found to be dominant in the MSC + G-CSF transplanted group and the BM transplanted group by week 4 post transplantation. In addition, we demonstrated that induction with G-CSF lead to CFU-GM colony formation from MSC compartment in vitro.

CONCLUSIONThese results indicate that under stimulation of G-CSF, mesenchymal derived stem like cells might differentiate into hematopoietic primitive stem cells in vivo and have the capacity to re-establish hematopoiesis in lethally irradiated mice. This study should provide an alternative transplantation treatment for malignancy.

Animals ; Bone Marrow Transplantation ; Cells, Cultured ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; administration & dosage ; Hematopoiesis ; drug effects ; radiation effects ; Hematopoietic Stem Cells ; cytology ; Male ; Mice ; Mice, Inbred BALB C ; Radiation Injuries, Experimental ; Radiation-Protective Agents ; administration & dosage ; Stem Cell Transplantation ; Whole-Body Irradiation

OBJECTIVETo study the characteristics of dermal mesenchymal stem cells (DMSC), and explore whether they could enhance hematopoiesis recovery in vivo as well as facilitate proliferation and differentiation of hematopoietic cells in vitro.

METHODSMultipotential stem cells from the murine dermal mesenchyme were dissociated and cultured as donor cells. After 2 approximately 3 passages, the growth status, cell cycle, immunophenotype and morphology of DMSC were analyzed. Hematopoietic cells were plated onto a feeder layer formed by DMSC, cell count and CFU-GM yields were observed dynamically. Female mice received 5 Gy (137)Cs radiation were injected with DMSC cultured for 2 - 3 passages via tail vein. Cell count and CFU-GM yields of the bone marrow were observed regularly. Pathological study of the liver, spleen and bone marrow was done to evaluate hematopoiesis recovery.

RESULTSMurine DMSC are adherent cells with a morphology of fibroblastoid and spindle and multiangle in shape. Immunophenotypes showed that CD(45), CD(34), HL-DR positive DMSC were 1 - 3%, CD(44) and CD(13) positive DMSC 75 approximately 95%. Cell cycle assay demonstrated 83% of DMSC being G(0)/G(1) phase. In vitro, the total cell count and CFU-GM yields in the experimental group were higher than those of the long-term culture bone marrow cells by the third week. The DMSC can sufficiently support the proliferation and differentiation of hematopoietic cells for seven weeks. In vivo, peripheral granulocytic count, cells in the bone marrow of one femoral bone and CFU-GM by the third week in the experimental group were much higher than those of controls. Genetic assay of the murine blood demonstrated Y chromosome.

CONCLUSIONThe DMSC have characteristics of stem cells. DMSC sped up hematopoiesis recovery of irradiated mice. DMSC as a feeder layer can support proliferation and differentiation of hematopoietic cells.

Animals ; Bone Marrow Transplantation ; Female ; Hematopoiesis ; Immunophenotyping ; Male ; Mesoderm ; cytology ; Mice ; Mice, Inbred BALB C ; Skin ; cytology ; Stem Cell Transplantation ; Stem Cells ; physiology

OBJECTIVETo establish a new and rapid GeXP based multiplex PCR assay for the detection and typing of human papillomavirus 6, 11, 31, 33 and 52.

METHODSNucleotide sequences of HPV6, HPV11, HPV31, HPV33 and HPV52 from NCBI were obtained and compared. Genotype-specific primers were then designed and the sensitivity and specificity of multiple PCR assay was evaluated. Optimized assay was further validated with 30 clinical specimens collected from the cervical secretions of patients.

RESULTSA GeXP based multiplex PCR was developed for sensitive detection and reliable differentiation of five HPV genotypes (HPV6, 11, 31, 33 and 52),

CONCLUSIONA GeXP based multiplex PCR assay is demonstrated to be a new and rapid technique for simultaneous detection and typing of 5 different human papillomaviruses.

Adult ; Female ; Humans ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods

OBJECTIVETo investigate the expression of thymosin beta10 (Tbeta10) and related changes of actin filament organization in human tumor cell lines with different metastatic potential.

METHODSFour groups of nine human tumor cell lines with different metastatic potential were analyzed for the expression of Tbeta10 mRNA detected by northern-blot and its peptide by immunohistochemical staining. The filamentous actin (F-actin) was stained with TRITC-phalloidin to detect changes in actin organization.

RESULTSIn comparison with the non and/or weakly metastatic counterparts, Tbeta10 was upregulated in highly metastatic human lung cancer, malignant melanoma and breast cancer cell lines. TRITC-phalloidin staining revealed less actin bundles and a fuzzy network of shorter filaments in the highly metastatic tumor cells, while in the non and/or weakly metastatic cancer cell lines, there were thick and orderly arranged actin filaments.

CONCLUSIONSTbeta10 levels correlate positively with the metastatic phenotype in human tumors currently examined. The increased metastatic potential of tumor cells is accompanied by the loss of F-actin and poorly organized actin skeleton. There is a consistent correlation between the elevated Tbeta10 expression and the disrupted actin skeleton.

Actins ; analysis ; Blotting, Northern ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Neoplasm Metastasis ; Thymosin ; analysis