Acupuncture Therapy ; adverse effects ; Humans

Bone Diseases ; complications ; pathology ; Bone Diseases, Infectious ; complications ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; complications ; microbiology ; Male ; Middle Aged ; Necrosis ; Osteomyelitis ; complications

AIMTo study the effects of cryopreservation length on the proliferative potential of hematopoietic cells derived from cord blood.

METHODSUsing Dextran-40 and 10% DMSO as cryoprotectants, separated nuclear cells were stored in liquid nitrogen after they were freezed according programme. One month or 4 months later, they were thawed and expanded in serum-free medium for culture and expansion of hematopoietic cell (SFEM) for 5 weeks. Dynamic results were detected every week.

RESULTSAt the 5th week of expanding, TNC were expanded for 1499.0 +/- 115.6-folds and 1513.0 +/- 110.4-folds, respectively. CD34+ cells and CFCs reached to their highest level at the 2nd week and at the 3rd week. CD34+ cells were expanded for 63.8 +/- 6.1-folds and 62.4 +/- 5.7-folds, respectively. CFCs were expanded for 53.8 +/- 6.3-folds and 54.8 +/- 6.7-folds, respectively. Between the two kinds of cells, statistical significant difference in proliferative potential wasn't detected.

CONCLUSIONIn ideal cryopreservative condition, the cryopreservation length would do not affect the proliferative potential of cord blood hematopoietic cells.

Cell Proliferation ; Cell Survival ; Cells, Cultured ; Cryopreservation ; methods ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Time Factors

To investigate the immune regulatory effects of human bone marrow mesenchymal stem cells on alloantigen T lymphocyte in vitro, human MSCs were isolated and expanded from bone marrow cells, and identified with cell morphology, and the phenotypes were assessed by immunohistochemistry and flow cytometry. As the stimulation factor of T lymphocytes proliferation, either PHA or dendritic cells isolated from cord blood were cocultured with CD2(+) T lymphocytes from peripheral blood mononuclear cells by magnetic beads with or without MSC in 96-well plats for seven days. T cell proliferation was assessed by [(3)H]-thymidine incorporation using a liquid scintillation counter. T cell subsets, Th1, Th2, Tc1 and Tc2 were analyzed by flow cytometry after co-culture of CD2(+) T cells with MSCs for 10 days. The results showed that a significant decrease of CD2(+) T cell proliferation was evident when MSC were added back to T cells stimulated by DC or PHA, and an increase of Th2 and Tc2 subsets were observed after co-culture of MSC with T lymphocytes. It is suggested that allogeneic MSC can suppress T cell proliferation in vitro and the cause of that was partly depend on interaction of cells and the alteration of T cell subsets.
Bone Marrow Cells ; cytology ; immunology ; CD2 Antigens ; immunology ; Cell Communication ; immunology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Flow Cytometry ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; cytology ; immunology ; T-Lymphocyte Subsets ; cytology ; immunology ; T-Lymphocytes ; cytology ; immunology

OBJECTIVETo discuss the reverse effect of Yinchenhao decoction(YCHD) in dimethyl nitrosamine (DMN)-induced hepatic fibrosis in rats.

METHODThe rat hepatic fibrosis model was established through the intraperitoneal injection with 1% dimethyl nitrosamine (DMN) with a dose of 1.0 mL x kg(-1) x d(-1) for consecutively three weeks, once for the first three days of each. The rats were randomly divided into six groups: the silymarin positive control group (50.0 mg x kg(-1) x d(-1), YCHD high (20.0 g x kg(-1) d(-1)), middle (8.0 g x kg(-1) x d(-1)) and low (3.2 g x kg(-1) x d(-1)) dose groups, the model group and the normal control group. The model group and the normal control group were orally administered with normal saline for consecutively five weeks. The pathologic changes in liver tissues were observed by HE staining. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), g-glutamyltransferase (g-GGT), hyaluronic acid (HA), laminin (LN), collagen type IV (CIV) and type III procollagen amino terminal peptide (PIIINP) in serum were determined. The metabolite profiling of amino acid and the content of hydroxyproline in liver tissues were also measured.

RESULTCompared with the model group, YCHD high and middle dose groups could significantly reverse the pathologic changes in liver tissues of rats. YCHD could reduce the levels of ALT, AST, gamma-GGT, HA, LN, CIV, PIIINP in serum and the content of hydroxyproline in liver tissues in a dose-dependent manner, and altered the metabolite profiling of amino acid in rat liver tissues.

CONCLUSIONYCHD has the effect in reversing dimethyl nitrosamine induced hepatic fibrosis in rats.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Collagen Type IV ; metabolism ; Dimethylnitrosamine ; adverse effects ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hydroxyproline ; metabolism ; Liver ; drug effects ; enzymology ; metabolism ; Liver Cirrhosis ; chemically induced ; drug therapy ; enzymology ; Male ; Rats ; Rats, Sprague-Dawley

To investigate the therapeutic effect of artesunate on mouse cytomegalovirus pneumonia, the BALB/c-nu mice were infected with murine cytomegalovirus-green fluorescent protein (MCMV-GFP) by nose dropping method. The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Guangzhou Medical University. The BALB/c-nu mice were randomly divided into five groups: control group, MCMV pneumonia group, and artesunate (60, 120, and 240 mg·kg^-1) groups. The survival rate, weights, and virus loads in lungs among the groups were observed. The degree of histopathologic changes in lungs was assessed directly by hematoxylin-eosin (HE) assay. MCMV-GFP expression was assessed by immunofluorescence. In addition, reverse transcription polymerase chain reaction (RT-PCR) analysis was performed to investigate the content of major immediate early 1 (Mie1) mRNA, and enzyme-linked immunosorbent assay (ELISA) was used to detect the changes of inflammatory factors, interleukin 10 (IL-10), IL-6, and tumor necrosis factor-α (TNF-α). Western blot analysis was used to detect the expression of the changes of nuclear factor-kappa B (NF-κB) signaling pathways in total proteins. Compared with MCMV group, artesunate (120 mg·kg^-1) significantly increased body weights of MCMV-infected nude mice over 30 days, and decreased the viral titer in lung homogenate, lung inflammation, and histological severity. Moreover, the administration of artesunate (120 mg·kg^-1) could downregulate the expression of phospho-NF-κB (p-NF-κB) p65 in the lungs of mice. The present study suggested that artesunate can protect the immunocompromised mice from MCMV-induced interstitial pneumonia via downregulating NF-κB signaling pathway, thus attenuating inflammation in the lungs.

OBJECTIVETo identify the type of the intermediate filament (IF) protein of Angiostrongylus cantonensis and analyze its tissue localization.

METHODSRecombinant pET-IF of antigen IF was expressed in E.coli with IPTG induction, and the expression products were purified by His.Bind column and identified for determining the type of the IF protein by Western blotting. Anti-IF antibody was prepared by multi-spot subcutaneous injection into mouse and used to detect the tissue slices of A. cantonensis by immunohistochemical analysis.

RESULTSThe antigen IF were correctly expressed and purified, and identified as a keratin located in the intestine wall and cytoplusma.

CONCLUSIONThe antigen IF is distributed in the intestine wall of A. cantonensis.

Angiostrongylus cantonensis ; cytology ; metabolism ; Animals ; Cell Nucleus ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Intermediate Filament Proteins ; classification ; genetics ; isolation & purification ; metabolism ; Protein Transport

OBJECTIVETo investigate the expressions of vascular endothelial growth factors (VEGF)-A, -C and -D and their prognostic significance and relation to angio- and lymphangiogenesis in gastric cancer.

METHODSThe expression of VEGF-A, -C and -D in 123 primary gastric cancers was detected by immunohistochemical staining. The lymphatic vessel density (LVD) and microvessel density (MVD) were assessed after immunohistochemical double-staining with D2-40 and CD34, respectively. The correlation between the expression of those VEGF factors and clinicopathological parameters were analyzed by univariate method. The overall survival was evaluated by Kaplan-Meier method and log-rank test. Multivariate analysis was carried out using Cox proportion hazard model.

RESULTSThe positive expression rate of VEGF-A, -C and -D in primary gastric cancer samples were 64.2%, 65.9% and 41.5%, respectively. High expression of VEGF-A, or -C or -D, or any two of them was correlated with high LVD (P < 0.05). High expression of both VEGF-A and -C was associated with high MVD, lymph node metastasis, LVI and MVI (P < 0.05). Both VEGF-C and -D high expression was correlated with LVI and lymph node metastasis (P < 0.05). The patients with high expression of these factors had a statistically shorter overall survival (P < 0.05). The patients with both VEGF-A and -C expression had the shortest survival (56 months). Multivariate analysis showed that VEGF-A high expression, MVD, lymph node metastasis and depth of tumor invasion were independent survival predictors (P = 0.033, 0.002, 0.019 and P < 0.001, respectively).

CONCLUSIONHigh expression of both VEGF-A and -C imply high potential of lymphangiogenesis, metastasis and poorer survival in gastric cancer patients. High expression of VEGF-C and -D may induce lymphangiogenesis and promote lymph node metastasis, but only VEGF-A is an independent predictor of survival.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Male ; Microvessels ; pathology ; Middle Aged ; Neovascularization, Pathologic ; Proportional Hazards Models ; Stomach Neoplasms ; metabolism ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism

OBJECTIVESTo explore a sensitive and specific method for detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma (DLBCL), and verify the credibility of the established method.

METHODSbcl-2/IgH hemi-nested PCR primers were designed using the professional primer design software. Fifty-two samples of pathologically diagnosed DLBCL and 10 fresh tonsil tissues were amplified using hemi-nested touch down-PCR to detect bcl-2/IgH gene rearrangement. The PCR products were cloned and sequenced.

RESULTSbcl-2/IgH gene rearrangement was detected in 6 of 52 DLBCL samples and 2 of 10 fresh tonsil tissues using one-way method. By using the hemi-nested PCR for the second round amplification, 5 of DLBCL were positive, but all of the fresh tonsil tissues were negative. The positive PCR products were sequenced and analyzed on the Internet, 3 of 8 cases obtained by one-way method were false positive, 5 positive cases amplified using hemi-nested PCR were all bcl-2/IgH gene rearrangement. PCR products of 3 false positive cases were homologous to BAC331191 and LLNLR-245D11 in human chromosome 19 and RP11-498P10 in chromosome 1.

CONCLUSIONThere are false positive results using common primers for detecting bcl-2/IgH gene rearrangement. The mechanism may be that highly homologous sequences to human genome exist in commonly used primers. The specificity of the diagnosis could be improved by hemi-nested PCR using the combination of primers we designed and the traditional ones.

Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Genes, bcl-2 ; genetics ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; Polymerase Chain Reaction ; methods

BACKGROUNDMarked interindividual variation exists in blood pressure response to benazepril, which is considered to have genetic basis. Our objectives were to evaluate whether the E112D polymorphism in the prolylcarboxypeptidase (PRCP) gene has impact on blood pressure response to benazepril.

METHODSHypertensive patients from Huoqiu County and Yuexi County of Anhui Province received daily treatment with an oral dosage of 10 mg benazepril for 15 days. Genotypes of the E112D polymorphism in the PRCP gene were determined by TaqMan SNP genotyping assay. Multivariate linear and Logistic regressions using generalized estimating equation model were performed in a total of 1092 patients to evaluate the association of PRCP genotypes and blood pressure response to benazepril.

RESULTSPatients carrying ED or DD genotype had a less systolic blood pressure reduction (adjusted beta = -3.7 + or - 1.1, P < 0.001), a less diastolic blood pressure reduction (adjusted beta = -3.1 + or - 0.8, P < 0.001) and a lower percentage of reaching target blood pressure defined as SBP lower than 140 mmHg and DBP lower than 90 mmHg (adjusted OR = 0.6, P = 0.005) than those patients carrying EE genotype. In addition, the results from stratified analysis by county (Huoqiu or Yuexi) were similar to those observed in the pooled population.

CONCLUSIONSOur data suggest that the E112D polymorphism in the PRCP gene may be a useful genetic marker to predict the antihypertensive effect of short-term benazepril treatment in hypertensive patients of Anhui Province, China.

Adult ; Aged ; Antihypertensive Agents ; therapeutic use ; Benzazepines ; therapeutic use ; Blood Pressure ; drug effects ; Carboxypeptidases ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Hypertension ; drug therapy ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; genetics ; physiology ; Young Adult