Carcinoma, Hepatocellular ; virology ; Hepatitis B virus ; genetics ; pathogenicity ; Humans ; Liver Neoplasms ; virology ; Receptors, Virus ; metabolism ; Virus Integration

OBJECTIVETo observe the correlation relationship between acupuncture at Dicang (ST 4), Hegu (LI 4) and Houxi (SI 3) on the affected side of peripheral facial paralysis patients and activated areas in brain functional areas and central regulation mechanism of acupuncture at Hegu (LI 4) treatment.

METHODSEighteen cases with left peripheral facial paralysis were randomly divided into a Hegu group, a Dicang group and a Houxi group, 6 cases in each group. They were treated with electroacupuncture at left Dicang (ST 4), Hegu (LI 4) and Houxi (SI 3), respectively, and were examined with fMRI covering the whole brain at the same time. The fMRI data was analyzed by SPM software.

RESULTSIt was found that the left precentral gyrus area and the left postcentral gyrus area were activated when electroacupuncture at left Hegu (LI 4), and the right precentral gyrus area and the bilateral postcentral gyrus area were activated when electroacupuncture at left Dicang (ST 4), and there was no activated area at precentral gyrus area and post central gyrus area when electroacupuncture at left Houxi (SI 3).

CONCLUSIONThe sensory importation information from Hegu (LI 4) and Dicang (ST 4) can converge and coincide in the brain and may influence each other.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Brain ; diagnostic imaging ; physiopathology ; Facial Paralysis ; diagnostic imaging ; physiopathology ; therapy ; Female ; Human Body ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography ; Young Adult

OBJECTIVETo develop a method for ng quantitation of circulating DNA in serum and explore the value in the diagnosis of cancer.

METHODSSerum DNA was extracted by commercial "genomic DNA extraction kit" and detected by fluorescent dye (SYBR green I) staining. Loss of heterozygosity (LOH) at BRCA1 (D17S579, D17S855) and p53 (TP53, D17S786) in serum DNA was analyzed by PCR-based method.

RESULTSSYBR green I dot staining could detect DNA as low as 2 ng. Using this method, we detected serum samples from 483 patients with various types of cancer and 150 healthy individuals. The mean DNA concentration in the normal controls was 22.2 +/- 13.4 ng/ml, while that in cancer patients was 81.3 +/- 98.3 ng/ml (P < 0.001). In 33 ovarian cancer patients with increased DNA level, 27(81.8%) displayed LOH in at least one of the four loci analyzed.

CONCLUSIONCirculating DNA in serum may become additional tumor marker for the diagnosis of cancer.

BRCA1 Protein ; genetics ; Biomarkers, Tumor ; blood ; DNA, Neoplasm ; blood ; genetics ; Female ; Genes, BRCA1 ; Genes, p53 ; Humans ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Neoplasms ; blood ; genetics ; Tumor Suppressor Protein p53 ; genetics

OBJECTIVETo investigate the antidepressant components of Polygala tenuifolia.

METHODThe chromatographic method was used to isolate and purify the chemical constituents, their structures were identified by spectral analysis, MTT method was applied to investigate their cytotoxic activities.

RESULTNine compounds were isolated from the roots of P. tenuifolia. Their structures were identified as sibiricose A, (1), sibiricose A5 (2), tenuifoliside A (3) and 3', 6-disinapoyl sucrose (4), sibiricose A6 (5), 3, 4, 5-trimethoxycinnamate (6), polygalaxanthone III (7), tenuifolioses A (8), tenuifolioses H (9) and some compounds' activities to PC12 were observed.

CONCLUSIONCompound 1 was isolated from this plant for the first time. Compounds 2,3 could protect PC12 cells damage induced by P. tenuifolia.

Animals ; Antidepressive Agents ; chemistry ; isolation & purification ; pharmacology ; Esters ; chemistry ; isolation & purification ; pharmacology ; Mice ; PC12 Cells ; Plant Roots ; chemistry ; Polygala ; chemistry ; Rats ; Sucrose ; chemistry

OBJECTIVETo evaluate the quantitative and functional changes of T helper (Th) cell subsets in the bone marrow of severe aplastic anemia (SAA) patients and the relationship between these changes and the patients hematopoietic function.

METHODSBy FACS, the quantity and ratio of Th1 and Th2 cells, the percentage of CD3(+)CD8(+) cells in the bone marrow were detected in 24 patients with SAA at active phase, 15 patients with SAA at recovery phase, and 16 normal controls. By radioimmunoassay, the serum levels of TNF-alpha, or IL-4 in 20 SAA patients at active phase, 12 at recovery phase and 16 normal controls were measured. The relationships between CD3(+)CD8(+) cells, TNF-alpha and Ret, ANC; and between Th1 cells and CD3(+)CD8(+) cells, TNF-alpha or Ret, ANC; between IL-4, balance of Th1/Th2 and Ret, ANC were evaluated.

RESULTSThe percentages of Th1 and Th2 cells, and ratio of Th1/Th2 in bone marrow of SAA patients at active phase were (4.87 +/- 2.64)%, (0.41 +/- 0.26)% and 21.22 +/- 5.07, respectively, being higher than those of normal controls [(0.42 +/- 0.30)% (P < 0.01), (0.24 +/- 0.17)% (P < 0.05) and (1.57 +/- 0.93) (P < 0.01), respectively] and all of them reduced to normal levels of SAA at recovery phase (P > 0.05). The percentage of CD3(+)CD8(+) cells significantly decreased from (32.32 +/- 8.69)% at active phase to (13.76 +/- 2.96)% at recovery phase (P < 0.01). The serum levels of TNF-alpha and IL-4 at active phase was (4.29 +/- 3.15) microg/L and (1.24 +/- 0.73) microg/L, respectively, being higher than those of normal controls (1.21 +/- 1.16) microg/L, (1.18 +/- 0.97) microg/L, but only the difference of TNF-alpha was statistically significant (P < 0.01). In recovery SAA patients, the serum levels of TNF-alpha significantly decreased to (1.46 +/- 1.41) microg/L (P < 0.01), and the levels of IL-4 increased markedly to (3.05 +/- 1.94) microg/L. The CD3(+)CD8(+) cells and TNF-alpha of patients negatively correlated with Ret (P < 0.05; P < 0.05) and ANC (P < 0.05; P < 0.05), Th1 cells correlated with CD3(+)CD8(+) cells and TNF-alpha positively (P < 0.01; P < 0.05), the Ret and ANC negatively (P < 0.01; P < 0.01), IL-4 and the balance of Th1/Th2 positively correlated with Ret and ANC (P < 0.05, P < 0.01; P < 0.01, P < 0.01).

CONCLUSIONThe bone marrow failure in SAA might be caused not only by the increase of Th1 cells, Th1 type effector cells and cytokines, but also by insufficient compensation of Th2 cells and Th2 type cytokines, which shifted the balance of Th1/Th2 favorable to Th1.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; pathology ; physiopathology ; Bone Marrow ; metabolism ; pathology ; CD3 Complex ; blood ; CD8 Antigens ; blood ; Child ; Female ; Hematopoietic System ; metabolism ; pathology ; physiopathology ; Humans ; Interleukin-4 ; blood ; Male ; Middle Aged ; Radioimmunoassay ; T-Lymphocytes, Helper-Inducer ; metabolism ; pathology ; Th1 Cells ; metabolism ; pathology ; Th2 Cells ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

OBJECTIVETo measure the subsets of dendritic cells 1 (DC1) in the bone marrow of severe aplastic anemia (SAA) patients and evaluate the relationships between the CD11c+CD83+ cells and Th1 cells, CD3+CD8+ cells or hematopoietic function and explore the role of DC1 in the pathogenesis of SAA.

METHODSBy FACS, the quantities and ratios of CD11c+CD1a+ cells, CD11c+CD83+ cells, Th1 cells, and CD3+CD8+ cells in the bone marrow of SAA patients and normal controls were detected respectively. The relationships between CD3+CD8+ cells and reticulocyte absolute value (Ret) or neutrophil absolute value (ANC), between Th1 cells and CD3+CD8+ cells, Ret or ANC, between CD11c+CD83+ cells, and Th1 cells, CD3+CD8+ cells, Ret or ANC were evaluated.

RESULTSIn normal controls' bone marrow, the percentages of Th1 cells, CD11c+CD1a+ cells, CD11c+CD83+ cells and the ratio of CD11c+CD83+/CD11c+CD1a+ were (0.42 +/- 0.30)%, (0.38 +/- 0.29)%, (0.37 +/- 0.32)% and 1.07 +/- 0.10, respectively. In untreated SAA patients, they were (4.87 +/- 0.54)%, (1.73 +/- 0.24)%, (3.38 +/- 0.56)% and 2.21 +/- 0.32 respectively, which were higher than that in normal controls (P < 0.01). In recovering SAA patients, the percentages of Th1 cells, CD11c+CD1a+ cells and CD11c+CD83+ cells decreased significantly to (0.53 +/- 0.22)%, (0.61 +/- 0.23)%, (0.65 +/- 0.22)%, respectively (P < 0.01). The ratio of CD11c+CD83+/CD11c+ CD1a+ in recovering SAA patients decreased to 1.37 +/- 0.25, which was similar to that in normal controls (P > 0.05). The percentage of CD3+CD8+ cells in untreated SAA patients was (32.32 +/- 10.22)%, and in recovering SAA patients decreased to (13.67 +/- 5.24)% (P < 0.01). The percentage of CD3+CD8+ cells in SAA patients was negatively correlated with their Ret and ANC (P < 0.05), while their Th1 cell percentages were positively correlated with their CD3+CD8+ cells (P < 0.01), and negatively correlated with their Ret and ANC (P < 0.01). SAA patient's CD11c+CD83+ cell percentages were positively correlated with their Th1 cell and CD3+CD8 cells (P < 0.01, P < 0.05), but negatively with their Ret and ANC (P < 0.01).

CONCLUSIONBoth immature DC1 and activated DC1 increased in the bone marrow of SAA patients, and the balance of DC1 subsets shifted from stable form to active one, which might promote Th0 cells to polarize to Th1 cells, and cause the over-function of T lymphocytes and hematopoiesis failure in SAA.

Adolescent ; Adult ; Anemia, Aplastic ; immunology ; Antigens, CD ; immunology ; Antigens, CD1 ; immunology ; Bone Marrow ; immunology ; CD11c Antigen ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Dendritic Cells ; immunology ; Female ; Humans ; Immunoglobulins ; immunology ; Male ; Membrane Glycoproteins ; immunology ; Th1 Cells ; immunology ; Young Adult

OBJECTIVETo study aberrant expression of cell cycle control genes in patients with myelodysplastic syndromes (MDS).

METHODSReverse transcription polymerase chain reaction (RT-PCR) was used to investigate the expression of cell cycle control genes (cyclin D2, cyclin D3, cyclin A1, cyclin E, CDK2, CDK4, CDK6, p21, p27, p57, Rb and E2F1) in bone marrow mononuclear cells (BMMNCs) from 29 normal control, 27 MDS and 19 de novo acute myeloid leukemia (AML).

RESULTSThe expression levels of cyclin D3 (0.65 +/- 0.17, P < 0.05) and cyclin A1 (0.48 +/- 0.04, P < 0.05) in MDS were higher than those in normal control and significantly lower than those in AML. The expression rates and levels of cyclin D2 (40.7% and 0.78 +/- 0.21) and cyclin E (51.9% and 0.52 +/- 0.10) in MDS were statistically higher than those in normal control and AML. The expression level of CDK2 in MDS (0.66 +/- 0.19, P < 0.01) was higher than that in normal control (0.42 +/- 0.04) and the expression rate of CDK6 in MDS (25.9%) higher than in normal control (3.4%, P < 0.05). There was no significant difference of the expression rates and levels of CDK4 in MDS, AML and normal control. The expression rates and levels of p21 (77.8% and 1.18 +/- 0.21) and p27 (48.1% and 1.14 +/- 0.40) in MDS were statistically higher than those in normal control and AML. The expression level of p57 in MDS (0.69 +/- 0.06) was higher than that (0.53 +/- 0.05, P < 0.01) in normal control but lower than in AML (0.96 +/- 0.16, P < 0.01). The expression rate (55.6%) and level (0.85 +/- 0.17) of Rb in MDS were significantly higher than those in normal control and AML. The expression rate (7.4%) and level (0.39 +/- 0.04) of E2F1 in MDS were comparable to those in normal control but lower than those in AML.

CONCLUSIONMDS clones have aberrant mechanism of cell cycle control: high expressions of cyclin family members, CDK2 and CDK6 may lead to high proliferation; high expression of p21 and p27 may cause the G1 phase arrest.

Adolescent ; Adult ; Cell Cycle Proteins ; genetics ; Child ; Cyclin-Dependent Kinase Inhibitor Proteins ; genetics ; Cyclin-Dependent Kinases ; genetics ; E2F1 Transcription Factor ; genetics ; Female ; Gene Expression Profiling ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; pathology ; Retinoblastoma Protein ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult

OBJECTIVETo study the role of changes of matrix metalloproteinase-2, 9 (MMP-2, 9) activity in the development of dimethylnitrosamine (DMN)-induced liver fibrosis in rats.

METHODSThe rat liver fibrosis model was established by peritoneal injection of DMN (at a dose of 10 mg/kg, 3 times a week, for 4 weeks). The dynamic changes of liver fibrosis were observed at different time points (1d, 2d, 3d, 1 week, 2 weeks, 4 weeks, 6 weeks and 8 weeks). The MMP-2, 9 activity was measured by zymogram method. Liver ultrastructure was observed by electron microscope. The expressions of type IV collagen (CIV), laminin (LN), type I collagen (CI) and alpha-smooth muscle actin (alpha-SMA) were examined by immunohistochemistry. The tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) content was measured by Western blot method.

RESULTSThe MMP-2, 9 activity (gray value) significantly increased in the 2d and 3d DMN model rats (2d: normal/model group, MMP-2: 54.72+/-4.56/70.76+/-7.63; F = 16.27, P < 0.05; MMP-9: 25.72+/-4.29/51.76+/-15.33, F=13.38, P < 0.05). The positive staining area percentage of CIV in the sinusoidal walls decreased in the 2d, 3d and 1 weeks model rats (2d: normal/model group, 6.06+/-1.35/2.86+/-0.63, F=69.12, P < 0.05), but significantly increased in the 4w model rats (normal/model group, 6.06+/-1.35/8.04+/-1.50, F=14.42, P < 0.05). There was a remarkable negative correlation between the MMP-9 activity and expression of CIV in the sinusoidal walls (r = -0.729, P < 0.05). Positive expressions of LN and CI increased, and the strongest positive staining of them displayed in the 4w model rats. The formation of basement membrane was also observed in the 4 weeks model rats. Expression of TIMP-2 significantly increased in the late stage of fibrosis.

CONCLUSIONSThe increase of MMPs activity, especially MMP-9 which degrades the CIV normally distributed under the sinusoidal endothelium is the important factor in the formation of sinusoidal capillarization. The deposition and reconstitution of LN and new synthetic CIV, adding the deposition of CI constitute the high density basement membrane. The increase of TIMP-2 expression in the late stage of the fibrosis may be one of reasons why natural resolution of DMN-induced liver fibrosis is difficult.

Animals ; Collagen Type IV ; analysis ; Laminin ; analysis ; Liver ; chemistry ; ultrastructure ; Liver Cirrhosis, Experimental ; enzymology ; pathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; analysis

OBJECTIVETo investigate the quantity, subset of dendritic cells (DC) and their costimulatory molecule expression in peripheral blood (PB) of the patients with myelodysplastic syndromes (MDS).

METHODSTotal DC (Lin1(+) HLA-DR(+)), myeloid DC (mDC) (Lin1(-) HLA-DR(+) CD11c(+)) and plasma DC (pDC) (Lin1(-) HLA-DR(+) CD123(+)) in fresh PB samples of 38 MDS patients and 19 normal controls were assayed by flow cytometry with the monoclonal antibodies. The expressions of costimulatory molecules CD80, CD86 and CD40 on these DCs were also assayed in the same way.

RESULTSThe number of total DC in PB of low-risk and high-risk MDS patients was significantly higher than that in normal controls [(33.7 +/- 7.0) x 10(6)/L, (56.3 +/- 29.0) x 10(6)/L vs (12.1 +/- 1.4) x 10(6)/L, respectively] (P < 0.05), that of mDC in PB of low-risk and high-risk MDS patients was higher than that of normal controls too [(16.7 +/- 6.3) x 10(6)/L, (28.7 +/- 17.6) x 10(6)/L vs (5.5 +/- 0.9) x 10(6)/L] (P < 0.05), but pDC in low-risk and high-risk MDS patients was not significantly higher than that in normal controls (P > 0.05). The percentage of total DC in PB mononuclear cells (PBMNC) of low-risk and high-risk MDS patients [(2.37 +/- 0.53)% and (3.58 +/- 1.39)% respectively and that of mDC (0.90 +/- 0.35)%, (1.51 +/- 0.70)% respectively] were higher than that of normal controls [(0.68 +/- 0.08)%, and (0.32 +/- 0.05)% respectively] (P < 0.05), but that of pDC in MDS cases was not higher than that of normal controls (P > 0.05). The expressions of CD80 and CD86 between MDS patients and normal controls had significant difference (P < 0.05).

CONCLUSIONSTotal DC and mDC were increased significantly in MDS, but pDC did not. The costimulatory molecules (CD80 and CD86) except CD40 expressed higher on the DC of MDS patients. It suggested that the inflammatory injury related APC increased in MDS, but the antitumour immunity related APC did not . What found here might be one of the mechanisms involved in the pathogenesis of MDS.

Adolescent ; Adult ; Aged ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Dendritic Cells ; immunology ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; metabolism ; pathology

OBJECTIVETo investigate the southern region of Zhejiang hepatitis E virus (HEV) infection.

METHODSA cluster sampling strategy was used to sample all blood donors from February to October in 2008 in Wenzhou blood center. Their blood was tested for IgG and IgM antibody against HEV. Reverse transcriptase-polymerase chain reaction(RT-PCR) and sequencing were applied to detect its genotype and sequence homology in HEV IgM-positive specimen.

RESULTSThe prevalence of anti-HEV IgG in 3044 cases of blood donors was 33.28%. IgG increased with age. There are certain increase in positive rates between the 20-year-old group and over 40 years of age group from 21.16% to 50.36%. The positive rate of IgM was 0.92%. The ratio of infection among different age group was the highest in the age range from 31 to 40 years and up to 1.90%. IgG and IgM through their negative and positive analysis of samples found in their group with donors age, sex and blood type does not significantly related to each other. Nucleic acids were found in three cases through PCR amplification in all 28 cases of HEV IgM positive samples. The total positive rate was one-thousandth, of which two cases for gene 4, 1 cases of infection for gene 1.

CONCLUSIONThe results indicate that there was a certain percentage of HEV virus in voluntary blood donors in south Zhejiang.

Adolescent ; Adult ; Blood Donors ; China ; epidemiology ; Female ; Genotype ; Hepatitis Antibodies ; blood ; Hepatitis E ; blood ; epidemiology ; virology ; Hepatitis E virus ; classification ; genetics ; immunology ; isolation & purification ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Male ; Middle Aged ; Phylogeny ; Young Adult