Objective: To investigate the role of human umbilical vein endothelial cells (HUVEC) in supporting hematopoiesis and the expansion of hematopoietic stem/progenitor cells in vitro. Methods: According to the fact that HUVEC supernatant has colony stimulating activity shown by methylcellulose colony-forming assay and HUVEC can maintain the survival of mononuclear cells for at least four weeks in vitro, CD34+ cells from umbilical cord blood were seeded with (HUVEC group) or without (control group) HUVEC monolayer. Every week cells were collected and counted, the frequency of CFU-GM was measured by using methylcellulose colony-forming assay, and the percentage of CD34+ and CD41a+ cells was measured by flow cytometry. Results: In control group，all the CD34+ cells died in two weeks. However, in HUVEC group,most nucleated cells and CD34+ cells were expanded by 68.1±14.8 fold and 6.6±1.4 fold，respectively at the third week while CFU-GM expansion reached its peak (5.7±2.1 fold) at the week 2. Moreover, the percentage of CD41a+ cells was enhanced significantly, reaching a maximum (15.6%) at the week 3. Conclusions:HUVEC can support hematopoiesis in vitro and expand the hematopoietic progenitor cells and CD41a+ cells in direct contact coculture.

OBJECTIVETo explore the relationship between Helicobacter pylori (Hp) infection and iron status using serum ferritin (SF) as a marker for total iron and to identify the related factors of iron nutritional status among preschool children.

METHODSBy cluster sampling, we recruited 475 preschool children aged 2 to 7 years. A structured questionnaire and diet form were sent to the parents of these children to obtain related information about the socioeconomic level and dietary intakes. After collecting blood samples, the following indexes were measured. Hp IgG antibodies were measured with a dot enzyme-linked immunoassay; hemoglobin, Hct, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width index (RDW) with automatic Complete Blood Count; SF with an immunoradiometric assay. Stool Hp antigen and occult bleeding were measured with ELISA among individuals who were Hp seropositive. Hp status was defined as positive when both serum and stool antigen tests were positive, Hp status was defined as negative when serum antigen test was negative; 24-hour weighting and recording methods were used to dietary survey for three days in May and December 2001, respectively, dietary intakes including energy, protein and micronutrient were calculated using nursery school nutrition software and evaluated by Chinese Dietary Reference Intakes (DRIs). Mann-Whitney test was used to compare mean ranks of SF in Hp-positive and Hp-negative children. To obtain an adjusted estimate of the impact of Hp infection on SF, a multivariate analysis of covariance was done to evaluate the different level of SF between Hp infected and non-infected status. The relationship between iron deficiency and gender, age, socioeconomic condition, iron intake, and calcium intake was assessed by univariate analysis. An unconditional multivariate logistic regression analysis was also performed. Iron deficiency status was dichotomized and placed as the dependent variable. Hp infection status was incorporated together with possible confounding factors as independent variables in a final logistic regression model. All the data were managed by EPI Info 5.01a and analyzed by SAS (Version 6.12).

RESULTSTotally 64 children were diagnosed as Hp-positive and 305 as Hp-negative. Mann-Whitney test and multivariate analysis of covariance both showed that SF concentration was significantly lower in Hp infected individuals than non-infected individuals. Adjusted mean level and 95% confidence interval of SF in infected and non-infected children was 23.62 microg/L (7.13 microg/L-78.26 microg/L), 33.48 microg/L (10.28 microg/L-109.06 microg/L), respectively. The relationship between Hp infection and iron deficiency status persisted in logistic regression analysis after adjusting for possible confounding factors (OR: 7.95; 95% CI 2.56 - 24.67).

CONCLUSIONIron nutritional status was reduced in Hp infected preschool children. Hp infection appears to be an independent risk factor or an added stressor on iron status among preschool children.

Antibodies, Viral ; blood ; Child ; Child, Preschool ; Erythrocyte Indices ; Female ; Ferritins ; blood ; Helicobacter Infections ; blood ; Helicobacter pylori ; immunology ; Hemoglobins ; analysis ; Humans ; Logistic Models ; Male ; Multivariate Analysis ; Nutrition Assessment

OBJECTIVETo explore the etiological factors of benign paroxysmal positional vertigo (BP-PV).

METHODSThe clinical data of 145 patients with BPPV were retrospectively reviewed. The impacts of gender and age on BPPV were analyzed. The relationship between the onset of BPPV and the internal ear ischemia was also explored.

RESULTSThe abnormality rate of auditory brainstem response (ABR) under high stimulus rate was 59.3% (86/145) in all the patients with, including 22.1% (32/145) in male and 37.2% (54/ 145) in female (P > 0.05). The abnormality rate of ABR under high stimulus rate were 37.9% (55/145) and 21.4% (31/145) in middle-aged (30-55 years) and old ( > 55 years) patients, respectively (P > 0.05).

CONCLUSIONThe onset of BPPV may relate to ischemic internal ear but is not relevant with gender and age.

Adult ; Age Factors ; Age of Onset ; Aged ; Evoked Potentials, Auditory, Brain Stem ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Time Factors ; Vertigo ; etiology ; pathology ; physiopathology ; Young Adult

Adolescent ; Humans ; Male ; Pulmonary Embolism ; etiology ; Sinus Thrombosis, Intracranial ; complications ; diagnosis

Objective:To explore the effect and underlying mechanism of koumine （Kou） at different concentrations （0， 100， 200， 400 μmol·L^-1） on the proliferation and apoptosis of colorectal cancer HCT-116 cells. Method:After 24 hours of in vitro intervention with HCT-116 cells by Kou， cell counting kit-8 （CCK-8） assay was used to detect its effect on cell proliferation. Flow cytometry was used to detect cell cycle， apoptosis， and reactive oxygen species （ROS） expression. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of forkhead box O3a （FoxO3a）. Cells were transfected with small interfering ribonucleic acid （siRNA）. Western blot was employed to detect the protein expression of the FoxO3a target gene. Result:Compared with the conditions in the blank group， Kou treatment reduced the proliferation rate of HCT-116 cells （P<0.05， P<0.01） in a dose-dependent manner， caused cell cycle arrest in the G₀/G₁ phase， and induced the apoptosis of HCT-116 cells （P<0.05， P<0.01）， which was positively correlated with the concentration of Kou. FoxO3a siRNA interference reduced the expression of FoxO3a and its downstream target genes cyclin-dependent kinase inhibitor 1A （p21）， cyclin-dependent kinase inhibitor 1B （p27）， and Bcl-2 interacting mediator of cell death （Bim） （P<0.01）. Kou treatment induced the activation of c-Jun N-terminal kinase （JNK） in HCT116 cells. SP600125 （JNK specific inhibitor） treatment inhibited the Kou-induced FoxO3a activation and the expression of its downstream target genes. N-acetyl cysteine （NAC） treatment reduced Kou-induced ROS levels （P<0.01） and JNK signal activation. The above results were significantly different from those in the blank group （P<0.01）. Conclusion:Kou can effectively inhibit the proliferation of HCT-116 cells and promote apoptosis， and the mechanism may be related to the regulation of the ROS/JNK/FoxO3a pathway.

OBJECTIVETo observe the influence of combined injection with human interferon (hlFNgamma) and human insulin-link growth factor-1 (hIGF-1) on regeneration and fibrosis of skeletal muscle after acute contusion.

METHODSA standard contusion model was reproduced at the right gastrocnemius in 64 male mice of 7 to 12 weeks. All the mice were randomly divided into 4 groups, such as group A (injection with hIFNgamma), group B (injection with hIGF-1), group C (injection with hIGF-1 and hIFNgamma), and group D (injection with physiological saline as control). All injections were introduced on day 10 after injury at local injured gastrocnemius. Before intervention (7 d following injury), and 4 d, 18 d, 32 d after intervention, the local injured gastrocnemius were harvested from 4 mice of each group. Then the expression of MHC- II b and vimentin was detected by real time PCR and immunohistochemistry techniques.

RESULTS(1) At the each time following intervention, the expression of MHC-II b mRNA and protein in local injured muscle of group B and C were significantly higher than those of group A and D. (2) After intervention,the expression of vimentin mRNA and protein in local injured muscle of group A, group B, and group C were more inhibited than those of group D. The inhibition of vimentin expression in group A and C was significant.

CONCLUSIONIt is indicated that injection of hIGF-1 into the injured skeletal muscle following acute contusion could enhance muscle regeneration,and inhibit fibrosis to some extent. (2) It is identified that hIFNgamma injected into injured muscle has the effect of anti-fibrosis, which is more significant than that of hIGF-1. (3) Combined injection with hIGF-1 and hIFNgamma could improve muscle regeneration and inhibit fibrosis simultaneously, and promote the healing of injured muscle.

Animals ; Immunohistochemistry ; Injections ; Insulin-Like Growth Factor I ; administration & dosage ; Interferon-gamma ; administration & dosage ; Male ; Mice ; Mice, Inbred C3H ; Muscle, Skeletal ; injuries ; Myosin Heavy Chains ; analysis ; genetics ; Polymerase Chain Reaction ; RNA, Messenger ; analysis ; Transforming Growth Factor beta1 ; analysis ; Vimentin ; analysis ; genetics ; Wound Healing ; drug effects

OBJECTIVETo investigate the relationship between serum alpha-fetoprotein (AFP) variant levels in patients with hepatocellular carcinoma (HCC) and cancer cells disseminating through blood.

METHODSSerum AFP variant levels were measured by crossed immunoaffino-electrophoresis in the presence of lectin before initial surgical treatment in HCC patients. Circulating tumor cells were simultaneously detected in pre-operative blood samples using reverse transcription-polymerase chain reaction (RT-PCR) for AFP mRNA.

RESULTSForty-six HCC patients with serum AFP positive were studied. Serum AFP variant level > or 20% was showed in 37 patients, among whom there were 22 (59.5%) showing AFP mRNA positive. In contrast, the positive AFP mRNA expression was only observed in 2 out of 9 patients (22.2%) with AFP variant level<20% (x(2)=4.02, P<0.05).

CONCLUSIONIn hepatocellular carcinoma patients, increased AFP variant levels are associated with a haematogenous spread of tumor cells.

Adult ; Aged ; Carcinoma, Hepatocellular ; blood ; Female ; Humans ; Liver Neoplasms ; blood ; Male ; Middle Aged ; RNA, Messenger ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; alpha-Fetoproteins ; analysis ; genetics

OBJECTIVETo observe the effects of the grafting of a platelet-derived growth factor gene-modified artificial composite skin on rat wounds with full thickness defect.

METHODSPlatelet derived growth factor-B (PDGF-B) eukaryotic expression plasmid was constructed, and the fibroblasts were transfected with it by liposome mediation. Artificial composite skins 1 and 2 were constructed respectively. The skin1 was composed of keratinocyte, porcine acellular dermal matrix and PDGF-B gene-transfected fibroblasts while the skin 2 contained keratinocyte, porcine acellular dermal matrix and fibroblasts. The two kinds of composite skin were grafted onto wounds on the rat back to form composite skin group 1 (C1) and 2 (C2), respectively, with 18 rats in each group. Eight rats with wounds without treatment served as control (C) group. The survival rate of the composite skin was observed at 2 post-operative weeks (POWs). The rat wounds were examined grossly on 2, 4 and 6 POWs for the calculation of wound contraction rate. Wound tissue samples were harvested for histological examination.

RESULTS(1) Up to 2 POWs, 14 grafts in C1 group survived completely, 3 with partial survival and 1 failure. In C2 group, 10 skin grafts survived completely, 4 with partial survival and 4 failures. (2) A scab was formed in the wound at 2 POW in C group. The surface of the grafted skin in C1 group was smooth, elastic, and showed good anti-friction properly, and it was better in quality compared with that in other two groups at 6 POW. (3) The wound contraction rate of the grafts in C group of rats was higher than that in C1 and C2 groups at 2, 4 and 6 POWs, while that in C1 was lower than that in C2 group. (4) Capillary formation was more intense in the grafted skins in C1 group at 2 POWs, and the epithelia differentiated well into 7 to 10 layers of epithelial cells with compact and orderly arrangement and evenly distributed fibrous tissue at 6 POWs.

CONCLUSIONRepair of the wound with artificial composite skin containing PDGF-B gene could improve the quality of wound healing.

Animals ; Cells, Cultured ; Female ; Humans ; Proto-Oncogene Proteins c-sis ; genetics ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; methods ; Skin, Artificial ; Swine ; Tissue Engineering ; Transfection ; Wound Healing

OBJECTIVETo establish a new method for the preparation of porcine acellular dermal matrix.

METHODSThe antigenicity of the porcine dermis was weakened by removing epidermal and dermal cells from the porcine skin through the digestion with low-concentration trypsin and repeated freeze-thaw cycles. Split thickness porcine skin was treated with 0.05% trypsin to remove the cells from the epidermis and dermis. Repeated freeze-thaw cycles were employed to further weed out the residual cells within the dermis. The prepared acellular dermis was then examined grossly, as well as histologically, and also by immunohistochemical method.

RESULTSNo cell could be identified in the prepared porcine acellular dermal matrix. The integral basement membrane was preserved on the surface of dermal matrix with compact dermal matrix collagen structure.

CONCLUSIONLow concentration trypsinization and repeated freeze-thaw cycles seemed to be a simple and effective method for the preparation of xenogeneic acellular dermal matrix.

Animals ; Dermis ; cytology ; transplantation ; Extracellular Matrix ; transplantation ; ultrastructure ; Freezing ; Skin Transplantation ; Swine ; Tissue Engineering ; methods ; Trypsin ; administration & dosage

A simple, rapid and sensitive colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) method was established to detect norovirus genotype GII. The method employed a set of six specially designed primers that recognized eight distinct sequences of RNA-dependant RNA polymerase and capsid protein gene for amplification of nucleic acid under isothermal conditions at 65 degrees C for 60 minutes. The amplification process of RT-LAMP was monitored by the addition of HNB (Hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by agarose electrophoresis. The specificity of the RT-LAMP was validated by detecting several different diarrhea viruses including norovirus genotype GII. The sensitivity was determined by serial dilutions of RNA molecules from in vitro transcription of norovirus genotype GII in parallel with conventional RT-PCR detection. The assay was further evaluated with 93 clinical specimens of diarrhea patients. The results showed that the sensitivity of RT-LAMP was 1 000 copies/microL with a high specificity and the relative sensitivity was at the same level as that of conventional RT-PCR. Positive rate of RT-LAMP in analysis of clinical specimens was approximately the same as that of conventional RT-PCR as well. This colorimetric RT-LAMP assay was potential for rapid detection of norovirus genotype GII on spot due to the observation of visual result with high specificity and sensitivity, time-saving and cost benefit.
Caliciviridae Infections ; diagnosis ; virology ; Colorimetry ; methods ; Feces ; virology ; Genotype ; Humans ; Norovirus ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods