Asthma ; diagnosis ; Child ; Humans ; Perception

AIM: To investigate the expression of matrix metalloproteinases (MMPs), membrane type MMPs (MT-MMPs) and their inhibitors (TIMPs) in human primary cultured prostatic cells and malignant prostate cell lines.METHODS: Reverse transcription-polymerase chain reaction-based measurements of the mRNA levels of MMP-2, MMP-7, MMP-9, MT1-MMP, MT3-MMP, TIMP-1 and TIMP-2 in relation to the house-keeping gene glyceraldehydes phosphate dehydrogenase were performed in cancerous and non-cancerous prostatic tissue samples, in primary cell cultures of epithelial cells, in both fibroblasts and smooth-muscle cells as stromal cells, and in the human malignant prostatic cell lines DU-145, LNCaP and PC-3.RESULTS: MMP-2 was mainly expressed in stromal cells. MMP-7 and MMP-9 showed their highest values in epithelial cells. MT1-MMP, MT3-MMP, TIMP-1 and TIMP-2 were found both in stromal and in epithelial cells, but there were some differences between the expression in fibroblasts and smooth-muscle cells. Different expression was also observed between the cells deriving from the primary cell cultures, the benign cell line BPH-1, and the malignant cell lines LNCaP, DU-145, and PC-3.CONCLUSION: These results concerning different expression of MMPs and TIMPs in cells from prostatic tissue suggest that a better insight into changes observed in prostatic tissue needs studies on cells cultured from the tissue.

OBJECTIVETo investigate the effect of hypoxia on the human pulmonary artery smooth muscle cells two pore domain potassium channels TASK-1 and the regulation of non-receptor tyrosine kinase c-Src in this process.

METHODSThe cultured human pulmonary artery smooth muscle cells (hPASMCs) were divided into: normal group, hypoxia 30 minute group, hypoxia 6 hours group and hypoxia 48 hour group, and hypoxia 48 hour + PP2 group, hypoxia 48 hour + PP3 group, hypoxia 48 hour + bpV group. Flow cytometry was used to analyze the cell cycle, RT-PCR and Western blot technique were carried out to detect the expression changes of TASK-1 mRNA and protein in different groups.

RESULTS(1) Cell Cycle Show: Compared with normal control group, with prolonged hypoxia, the percentages of hPASMCs in S phases of cell cycle were increased. While compared with hypoxia 48 hour group, the percentages of hypoxia 48 hour + PP2 group hPASMCs in S phases of cell cycle were decreased. The expression of TASK-1 mRNA on hPASMCs in acute hypoxia 6 hour group was increased, while the expression of TASK-1 protein on hPASMCs in the acute and chronic hypoxia group was decreased, and the expression of TASK-1 mRNA on hPASMCs in the chronic hypoxia group was decreased; After pre-incubation of a potent and selective inhibitor of the Src family of protein tyrosine kinases PP2, the expression of TASK-1 mRNA and protein in hypoxia 48 hour group was increased, however after pre-incubation of the inhibitor of the Src family of protein tyrosine phosphatase bpV, the expression of TASK-1 protein in hypoxia 48 hour group was decreased.

CONCLUSIONHypoxia promotes human pulmonary artery smooth muscle cell proliferation, and non-receptor tyrosine kinase c-Src may participate in the expression of two pore domain potassium channels TASK-1 regulated by hypoxia. Therefore, we hypothesized that TASK-1 channels and c-Src participatein the acute and chronic hypoxic human pulmonary vasoconstriction.

Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Humans ; Myocytes, Smooth Muscle ; cytology ; Nerve Tissue Proteins ; metabolism ; Potassium Channels, Tandem Pore Domain ; metabolism ; Pulmonary Artery ; cytology ; RNA, Messenger ; Vasoconstriction ; src-Family Kinases ; metabolism

OBJECTIVEPrepare konjac glucomannan-hydroxypropyl methyl cellulose (HPMC) compression coated tablets and study the effects of the formulation, technics and in vitro dissolution condition on drug release behavior to elevate the colon-specific effects of preparation.

METHODBerberine hydrochloride core tablets were prepared by wet granulation technique and konjac glucomannan-HPMC mixture as the coating layer were used with compression coated technique. The effects of the formulation and technics on drug release behavior were investigated by dissolution test. The erosion of coat layer during dissolution test was investigated.

RESULTDrug almost not released in dissolution medium stimulating gastric and intestinal condition, and released completely by coating layer erosion and rupture by enzyme in stimulating colonic condition. Drug release decreased with decreasing the ratio of konjac glucomannan-HPMC and increasing coat weight (P < 0.05), compression force was not found to be a significant factor on drug release. Drug release increased with increasing the concentration of beta-mannase in dissolution medium (P < 0.05), rotation speed has no effect on drug release. The release of drug was correlative with erosion of coat layer. The mechanism of drug release were diffusion and erosion.

CONCLUSIONThe konjac glucomannan-HPMC compression coated tablets was a promising delivery system for drugs to be delivered to the colon.

Administration, Oral ; Amorphophallus ; chemistry ; Berberine ; administration & dosage ; chemistry ; pharmacokinetics ; Colon ; metabolism ; Drug Compounding ; methods ; Drug Delivery Systems ; Hypromellose Derivatives ; Mannans ; chemistry ; isolation & purification ; Methylcellulose ; analogs & derivatives ; chemistry ; Plants, Medicinal ; chemistry ; Tablets, Enteric-Coated

Objective To investigate homografting vascular endothelial progenitor cells(EPCs)for preventing restenosis formation of carotid artery in New Zealand white rabbit models.Methods EPCs of New Zealand white rabbits were isolated,confirmed and expanded though the injured carotid arterial endothelium of rabbit model induced by dilatation with a 2.5 F balloon;and then EPCs were transplanted into the injured endothelium of the cells transplantation group(n=13,3 of them were transplanted with fluorencently-labeled- EPCs),while equal volume of saline without EPCs was injected into the injured endothelium in the control group(n=8).Histopathology was performed at 4 days after transplantation for the 2 rabbits,with fluorencently-labeled-EPCs.All of the rest remained rabbits were killed 4 weeks later for histological examinations.Results The histopathological slides showed that the fluorescence-positive expression existed in the injured endothelium 4 days after transplantation.At 4 weeks after the EPCs transplantation,there were less restenosis and less vascular wall thickening in the rabbits of cells transplantation group than those of the control group(P＜0.01).Conclusion The local interventional homografting heterogeneous endothelial progenitor cells can prevent restenosis after the carotid artery angioplasty in New Zealand White rabbit model. (J Intervent Radiol,2007,16:95-98)

BACKGROUNDActinomycosis is a rare indolent infectious disease caused by Actinomyces. Although pulmonary actinomycosis is thought to be more prevalent in developing countries, data from developing countries are scanty. This study was to reveal the current situation of pulmonary actinomycosis in developing countries and the difference from that in developed countries.

METHODSPatients fulfilling the inclusion criteria for pulmonary actinomycosis from Peking Union Medical College Hospital in China between January 2003 and December 2014 were retrospectively analyzed. Baseline characteristics, clinical symptoms, underlying diseases, diagnostic methods, pulmonary function test results, chest computed tomography (CT) tests, fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) tests, initial diagnosis, treatment and prognosis were retrieved from medical records and analyzed.

RESULTSTwenty-six patients were included in this study (mean age 52.0 + 13.1 years). The ratio of male to female was 1.17:1. Most common clinical symptoms were cough (15/26), sputum (12/26) and hemoptysis (12/26). Chest CT findings presented as masses (13/26), nodules (10/26) and infiltrates (3/26). FDG-PET had an increased standardized uptake value and 4/6 patients were misdiagnosed as malignancy. Many kinds of antibiotics were used in the treatment of pulmonary actonomycosis and all got favorable results. Five patients receiving complete resection of the lesion were cured without postoperative use of antibiotic.

CONCLUSIONSPulmonary actinomycosis is a rare disease even in developing countries, and both misdiagnosis and missed diagnosis are common. FDG-PET seems useless in the differential diagnosis, and complete resection of the pulmonary lesion without postoperative antibiotic therapy might be enough to achieve cure.

Actinomycosis ; diagnosis ; diagnostic imaging ; metabolism ; Adult ; Aged ; China ; Female ; Fluorodeoxyglucose F18 ; Humans ; Lung Diseases ; diagnosis ; diagnostic imaging ; metabolism ; Male ; Middle Aged ; Positron-Emission Tomography ; Radiography ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo review the research progress on Type IV secretion system (T4SS) in Helicobacter pylori.

DATA SOURCESThe data used in this review were identified by searching of PUBMED (1995 - 2007) online resources using the key terms 'Type IV secretion system' and 'Helicobacter pylori'.

STUDY SELECTIONMainly original articles and critical reviews written by major pioneer investigators of this field were selected.

RESULTSThe research progress on T4SS in Helicobacter pylori was summarized. The structure and function was discussed.

CONCLUSIONST4SS is not only involved in toxin secretion and injection of virulence factors into eukaryotic host target cells, but also involved in horizontal DNA transfer to other bacteria and eukaryotic cells, through DNA uptake from or release into the extracellular milieu. It provides a new insight into the pathogenicity of Helicobacter pylori and a novel target for antimicrobials development. However, many challenges remain for us in understanding the biological role of T4SS in Helicobacter pylori.

Bacterial Proteins ; metabolism ; DNA-Binding Proteins ; Gene Transfer, Horizontal ; Helicobacter pylori ; genetics ; metabolism ; pathogenicity ; Multigene Family

To compare the growth characteristics of non-hematopoietic adult stem cells (NASC) derived from rat fetal blood and rat bone marrow in vitro, and to study the differentiation of these stem cells into neuron-like cells in vitro, the fetal blood of pregnant rats and bone marrow of adult rats were sterilely collected; mononuclear cells (MNC) were isolated by using standard Ficoll-hypague techniques and then cultured in DMEM/LG containing 10% fetal bovine serum (FBS). The acquired NASCs were subcultured for passage. The immunophenotype of NASCs was detected by flow cytometry. The expanded NASCs were induced to differentiate into neurons-like cells by beta-mercaptoethanol (beta-ME), dimethylsulfoxide (DMSO), butylated hydroxyanisole (BHA). The specific markers of these neuron-like cells were detected by immunocytochemistry. The results showed that two kinds of subcultured NASCs showed homogeneous spindle-shaped and expressed antigens CD44 and CD54, but did not expressed CD11b and CD45. The both induced cells were similar to neuron in morphology and were positive for nestin and neuron-specific enolase (NSE), but negative for glial fibrillary acidic protein (GFAP). It is concluded that no significant difference of NASCs derived from pregnant rat fetal blood and adult rat bone marrow found in cell morphology and biological characteristics. NASCs of both origins can be induced to differentiate into neuron-like cells, so fetal blood can be regarded as another resource of NASC.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; physiology ; Female ; Fetal Blood ; cytology ; Male ; Multipotent Stem Cells ; cytology ; physiology ; Neurons ; cytology ; Pregnancy ; Rats ; Rats, Wistar ; Stem Cells ; cytology

OBJECTIVETo establish a new method based DNA chip technique for detecting HCV genotypes.

METHODSGenotyping probes were designed according to the sequence of HCV 5' NCR to generate DNA chip. The probes on DNA chip contains 5 major genotypes and 8 subtypes. The DNA fragment amplified by labeling Cy5 fluorescence was hybridized with DNA chip.

RESULTSFifty-five out of 65 isolates detected by DNA chip belonged to 1b- DNA sequencing of form a part of the isolates was used as the control. The results of both were completely consistent.

CONCLUSIONThe method is simple and rapid with high specificity and sensitivity. It can be applied in detection of HCV RNA and genotypes.

5' Untranslated Regions ; genetics ; Genotype ; Hepacivirus ; classification ; genetics ; Humans ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis ; methods ; Polymerase Chain Reaction ; RNA, Viral ; blood ; Sensitivity and Specificity

OBJECTIVEMesenchymal stem cells (MSCs) from bone marrow are capable of differentiating into cells of different tissue lineages such as bone, cartilage, and adipose tissue and are the best candidates for tissue engineering. It is well accepted that umbilical cord blood (UCB) is a source for hematopoietic stem cells. However, controversy exists as to whether UCB contains MSCs and can serve as a source of MSCs. Therefore, the aim of this study was to explore the biological characteristics and inducing differentiation ability of in vitro expanded UCB MSCs.

METHODSUCB was collected on normal full term delivery of infants with informed consent (n = 35) obtained from the mothers. Mononuclear cells (MNCs) were isolated from UCB by gravity centrifugation and cultured with DMEM including 10% fetal bovine serum. The morphology was observed under microscope per day. Cytochemical staining was carried out and flow cytometry was used to examine the surface antigen phenotype. Fifth passage cells were transferred into a different medium and osteogenic differentiation, adipogenic differentiation, and neurogenic differentiation were assessed.

RESULTSMSCs could be isolated and cultured from MNCs of a few UCB sources. These cells displayed fibroblast-like morphology. They withstood over 20 passages without significant structural changes. These MSCs were negative for alkaline phosphatase (ALP) staining and positive for alpha-naphthol butyric acid esterase (NBE) staining. Expression of CD(29), CD(44)and CD(105), especially the human MSCs-specific markers SH-2 and SH-3 were observed, but CD(3), CD(14), CD(19), CD(34) and CD(45) could not be found, indicating that these cells were not of hematopoietic origin. Exposure of these MSCs to serum-free osteogenic condition, they could differentiate into bone cells and form mineralized matrix as evidenced by Alizarin red staining 2 weeks later. When these UCB-derived MSCs were cultured in adipogenic medium, morphologic changes in cells as well as the formation of neutral lipid vacuoles were noticeable as early as 1 week after induction and visualized by staining with oil-red O. Surprisingly, these MSCs were also able to differentiate into neuroglial-like cells. Morphology of these induced cells resembled that of neurons. Immunocytochemistry showed that they expressed Nestin and neuron-specific enolase (NSE), but not glial fibrillary acidic protein (GFAP).

CONCLUSIONUCB does contain MSCs. These MSCs, which are multipotent, could be isolated and cultured from a few UCB sources. UCB might serve as an alternative source of MSCs to bone marrow.

Alkaline Phosphatase ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Proliferation ; Centrifugation, Density Gradient ; Culture Media, Conditioned ; Fetal Blood ; cytology ; Flow Cytometry ; Histocytochemistry ; Humans ; Infant, Newborn ; Mesenchymal Stromal Cells ; metabolism ; Phosphopyruvate Hydratase ; metabolism