OBJECTIVEA study was conducted to investigate the effects of Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) on the expression of regulated upon activation normal T-cell expressed and secreted (RANTES) and fractalkine in human umbilical vein endothelial cells (HUVECs).

METHODSHUVECs were incubated with different concentrations of Pg-LPS (200, 500, and 1000 ng x mL(-1)) for 1, 6, 12, and 24 h, respectively. Then real time quantitative polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent method (ELISA) were adopted to detect the protein levels and mRNA levels of RANTES and fractalkine.

RESULTSThe RANTES protein levels and mRNA levels, as well as fractalkine mRNA levels, were significantly higher in all experimental groups of 1, 6, and 12 h than in the control group (P<0.05), except the expression of RANTES mRNA in 200 ng x mL(-1) group of 12 h and RANTES protein in 200 ng x mL(-1) group of 1 h. The expression levels of RANTES mRNA and fractalkine mRNA were highest in 1000 ng x mL(-1) group of 6 h and were 4.88- and 6.20-fold higher, respectively, than those in the control group. The expression levels of RANTES protein, mRNA, and fractalkine mRNA decreased 6 h after stimulation, and were significantly higher than those in the control group (P<0.05) in the RANTES and fractalkine in HUVEC, and such expression is important in the development of atherosclerosis 500 ng x mL(-1) group of 24 h. There was a significant difference between the expression of fractalkine mRNA in 1000 ng x mL(-1) group of 6 and 12 h than in the control group (P<0.05).

CONCLUSIONPg-LPS infection might up-regulate the expression of RANTES and fractalkine in HUVEC, and such expression is important in the development of atherosclerosis.

Atherosclerosis ; Cells, Cultured ; Chemokine CCL5 ; genetics ; metabolism ; Chemokine CX3CL1 ; analysis ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lipopolysaccharides ; pharmacology ; Porphyromonas gingivalis ; immunology ; isolation & purification ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Up-Regulation

Aquilaria agallocha can produce fragrant agarwood used for incense, traditional medicine and other products. An efficient plant regeneration system was established via organogenesis from shoots developed from seedlings of Aquilaria agallocha. Shoots generated many buds on MS medium supplemented with 1.3 micromol/L BA (6-benzylaminopurine) in the first 7 weeks, and the buds elongated on MS medium with 1.3 micromol/L BA+0.5 micromol/L NAA (naphthaleneacetic acid) in another 7 weeks, 2.3 shoots 2 cm in length per explant were obtained within 14 weeks. Plantlets were rooted on 1/2 MS medium after being immersed in 5 micromol/L NAA for 48 h, 96.7% of the roots grew up two weeks later. All plantlets that survived acclimatization grew well in the pots.
Agriculture ; methods ; Cell Culture Techniques ; methods ; Forestry ; methods ; Phytotherapy ; Plant Growth Regulators ; metabolism ; Plant Shoots ; drug effects ; growth & development ; Survival Analysis ; Survival Rate ; Thymelaeaceae ; drug effects ; growth & development

To explore influence of platelet donation on donor's megakaryocytopoiesis, platelet counts and plasma concentrations of thrombopoietin (TPO), interleukin-3 (IL-3), IL-6 and nitric oxide (NO) were determined in 42 frequent platelet donors (undergoing plateletpheresis more than once a month for 24 months and their mean platelet yield of collection was 2.5 x 10(11)), in 62 limited platelet donors (undergoing plateletpheresis less than once a month for 24 months) after a donation-free period of > 5 weeks and in 40 whole blood donors who never undergoing plateletpheresis after a donation-free period of > 6 months. The results showed that the TPO levels was significantly lower in frequent platelet donors than in limited platelet donors (P < 0.01) and whole blood donors (P < 0.01). There were no significant differences between three groups in platelet counts, IL-3, IL-6 and NO. These findings suggest that the number of megakarocytes significantly increased in frequent platelet donors.
Adult ; Blood Donors ; Humans ; Interleukin-3 ; blood ; Interleukin-6 ; blood ; Megakaryocytes ; cytology ; Platelet Count ; Plateletpheresis ; adverse effects ; Thrombopoiesis ; Thrombopoietin ; blood

Objective To investigate the effects of autologous bone mesenchymal stem cells(MSC) transplantation on malignant arrhythmia induced by electrophysiological (EP) stimulation and cardiomyocyte ion channels remodeling in a mini-swine model of acute myocardial infarction (AMI). Methods Immediately after AMI(LAD occluded for 120 min),MSC(10×107,labeled by colloidal gold and cocultivated with 5-azacytidiRe,5-aga,n=12) or equal volume saline (n=10) were iniected through over-the-wire (OTW) balloon in LAD at distal over D1.EP stimulation is performed after 2 hours and 4 weeks in both groups to induce arrhythmia.The variance of heterogeneity of sodium currents(INa)and INa steady-state inactivation curves in different zones of infracted wall were investigated by patch clamp technology and the relationship between ionic channel and ventrieular arrhythmia is analyzed.Results EP induced malignant ventficular arrhythmia (VT) rate was similar (MSC 75% vs.saline 90%.P=0.455) at 2 hours post AMI and was significantly lower in MSC group(25% vs.80%,P=0.012) at 4 weeks post AMI.The Peak INa current densities of the Endo,Media and Epi were significantly lower in MSC group[(-14.04±3.82)pA/pF,(-29.26±5.70)pA/pF,(-12.43±3.04)pA/pF] compared those in saline group [(-9.71±3.38)pA/pF,(-18.98 ±4.05)pA/pF,(-8.47±3.34)pA/pF,all P<0.05].The IHa steady-state inactivation curves of the Epi,Endo and Media in mini-swine with VT in MSC group [(-126.2±10.9)mV,(-106.7±11.9)mV,(-105.4±11.0)mV] were similar as those in saline group with VT[(-129.1±10.9)mV,(-112.2±9.9)mV,(-109.7±9.2)mV,all P>0.05] while significantlylower compared to MSC group without VT[(-93.1±13.8)mV,(-95.2±15.5)mV,(-103.4±8.7)mV,all P<0.05].The multiple logistic regression analysis showed that INa current density (RR=1.449,95%CI 1.276-2.079,P=0.029) and INa steady-state inaetivation curves(RR=1.092,95%CI 1.008-1.917.P=0.012) were the independent factors for reduced VT.Conclusions Autologous MSC attenuated malignant ventricudar arrhythmia induced by EP at 4 weeks in nlini-swine with AMl which might due to altered eardiomyocyte ion channels remedcling induced by MSC.

The ZM-1 tissue microarrayer designed by our groups is manufactured in stainless steel and brass and contains many features that make TMA (tissue microarray) paraffin blocks construction faster and more convenient. By means of ZM-1 tissue microarrayer, biopsy needles are used to punch the donor tissue specimens respectively. All the needles with the punched specimen cylinders are arrayed into the array-board, with an array of small holes dug to fit the needles. All the specimen cylinders arraying and the TMA paraffin block shaping are finished in only one step so that the specimen cylinders and the paraffin of the TMA block can very easily be incorporated and the recipient paraffin blocks need not be made in advance, and the paraffin used is the same as that for conventional pathology purpose. ZM-1 tissue microarrayer is easy to be manufactured, does not need any precision location system, and so is much cheaper than the currently used instrument. Our method's relatively cheap and simple ZM-1 tissue microarrayer technique of constructing TMA paraffin block may facilitate popularization of the TMA technology.
Biopsy, Needle ; instrumentation ; Equipment Design ; Female ; Humans ; Immunohistochemistry ; Male ; Neoplasms ; enzymology ; Paraffin ; Tissue Array Analysis ; instrumentation

BACKGROUNDMiddle mediastinal masses comprise a wide variety of tumors but may also reflect lymphadenopathy, and thus remain an interesting diagnostic challenge. We performed positron emission tomography (PET) of mediastinal masses in order to evaluate the ability of PET to predict the malignancy of these tumors. We compared histologic findings, videomediastinoscopy, computed tomography (CT), and PET-CT in patients with mediastinal disease.

METHODSThirty-two patients were evaluated with CT, PET-CT and videomediastionoscopy, and all studies were performed within four weeks in each patient. (11)C-choline as a PET tracer was used to visualize masses. PET data were evaluated using the standardized uptake value (SUV) and were compared with pathologic data.

RESULTSThere were 13 men and 19 women aged from 21 to 74 (mean 45.2) years. Among the patients with mediastinal diseases, sarcoidosis was diagnosed in 12 patients, tuberculosis in 5 patients, lymphoma in 5 patients, and noncaseating granulomata without classical "sarcoid" finding in 3 patients. N2 or N3 nodal metastasis was revealed in 6 of 7 patients who had non-small cell lung cancer or suspected lung cancer, and one was negative (the pathological diagnosis was reactive hyperplasia). The accuracies for correctly diagnosing mediastinal masses for CT, PET-CT and videomediastinoscopy were 38% (12/32), 63% (20/32), and 91% (29/32) respectively. The diagnostic accuracy of videomediastinoscopy was superior to that of PET-CT (chi(2) = 11.130, P < 0.001). The SUVs were similar among these diseases. On the other hand, if the diagnostic classification was benign vs malignancy, the accuracies for CT, PET-CT and videomediastinoscopy were 53% (17/32), 75% (24/32), 100% (32/32) respectively. The diagnostic accuracy of videomediastinoscopy was superior to that of PET-CT (chi(2) = 22.042, P < 0.001). The SUV of malignant lesions (6.9, 3.2 - 9.8; n = 11) appeared to be higher than that of benign lesions (4.9, 2.9 - 8.3; n = 21), however, this difference was not statistically significant (P = 0.054).

CONCLUSIONSTo diagnose lesions located in the middle mediastinum, videomediastinoscopy possesses the highest diagnostic accuracy, and therefore remains the gold standard. PET-CT is valuable for differential diagnosis of benign vs malignant lesions, CT alone or PET alone (SUV) may provide misdiagnosis in a substantial proportion of patients with mediastinal masses.

Adult ; Aged ; Carbon Radioisotopes ; Female ; Humans ; Male ; Mediastinal Diseases ; diagnosis ; Mediastinoscopy ; methods ; Middle Aged ; Positron-Emission Tomography ; Tomography, X-Ray Computed ; Video Recording

OBJECTIVETo explore the response of rat bone marrow mesenchymal stem cells (MSCs and calvarial osteoblasts to mechanical strain and the consequent changes of cytoskeleton F-actin.

METHODSBone marrow MSCs and calvarial osteoblasts were isolated from SD rats and cultured in vitro. Mechanical stretch was performed on passage 3 cells at 2 000 microepsilon for 0, 2, 6 and 12 hours using four-point bending system. The response of cells and the distribution of F-actin were observed using fluorescent staining under laser scanning confocal microscope and the morphological parameters were quantified using image analysis software Laserpix.

RESULTSUnder mechanical stretch, the fluorescent staining decreased obviously at both MSCs and osteoblasts, and F-actin filaments were rearranged and became tenuous, thinner, and abnormally distributed. The outline of nucleus became unclear and apoptotic changes were observed at some of both cells. Cellular size decreased more significantly in MSCs than in osteoblasts. Quantity analysis showed that total area of cells, total fluorescent density and green fluorescent density (F-actin) were all significantly decreased in MSCs (P < 0.05 or P < 0.01), and total fluorescent density, green fluorescent density and red fluorescent density (nuclei) did also in osteoblasts (P < 0.05 or P < 0.01).

CONCLUSIONMechanical stretch caused extensive response on both MSCs and osteoblasts which led to the rearrangement of F-actin filament and apoptosis in some of these cells. MSCs were more sensitive to mechanical strain than osteoblasts.

Actin Cytoskeleton ; metabolism ; Actins ; metabolism ; Animals ; Bone Marrow Cells ; Cells, Cultured ; Cytoskeleton ; Mesenchymal Stromal Cells ; Microtubules ; Osteoblasts ; Rats ; Stress, Mechanical

OBJECTIVETo study the clinicopathologic features, diagnosis and differential diagnosis of malignancies in giant cell tumor (MGCT).

METHODSThe clinicopathologic features of 13 cases of MGCT were retrospectively reviewed.

RESULTSThirteen cases of MGCT were found amongst a total of 603 cases of giant cell tumor encountered. Six of the 13 cases represented concurrent malignancy in giant cell tumor while the remaining 7 cases was malignant transformation in recurrent giant cell tumor. The age of the patients ranged from 21 to 71 years (mean age = 39.5 years) in the first group and from 27 to 52 years (mean age = 36.7 years) in the second group. In concurrent MGCT, a high-grade sarcoma component was present in conjunction with the giant cell tumor component. In malignant transformation of recurrent giant cell tumor, the original tumor was giant cell tumor and the recurrence showed features reminiscent of malignant fibrous histiocytoma.

CONCLUSIONSThe diagnosis of malignancies in giant cell tumor requires correlation of clinical, radiologic and pathologic features. The entities need to be distinguished from other giant cell-rich tumors including primary malignant fibrous histiocytoma and giant cell osteosarcoma.

Adult ; Aged ; Bone Neoplasms ; diagnostic imaging ; pathology ; surgery ; Cell Transformation, Neoplastic ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Giant Cell Tumor of Bone ; diagnostic imaging ; pathology ; surgery ; Histiocytoma, Malignant Fibrous ; pathology ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasms, Second Primary ; pathology ; Osteosarcoma ; pathology ; Radiography ; Sarcoma ; pathology ; Young Adult

AIMUnderstanding the response of mesenchymal stem cells (MSCs) to mechanical strain and their consequent gene expression patterns will broaden our knowledge of the mechanobiology of distraction osteogenesis.

METHODOLOGYIn this study, a single period of cyclic mechanical stretch (0.5 Hz, 2,000 microepsilon) was performed on rat bone marrow MSCs. Cellular proliferation and alkaline phosphatase (ALP) activity was examined. The mRNA expression of six bone-related genes (Ets-1, bFGF, IGF-II, TGF-beta, Cbfa1 and ALP) was detected using real-time quantitative RT-PCR.

RESULTSThe results showed that mechanical strain can promote MSCs proliferation, increase ALP activity, and up-regulate the expression of these genes. A significant increase in Ets-1 expression was detected immediately after mechanical stimulation, but Cbfa1 expression became elevated later. The temporal expression pattern of ALP coincided perfectly with Cbfa1.

CONCLUSIONThe results of this study suggest that mechanical strain may act as a stimulator to induce differentiation of MSCs into osteoblasts, and that these bone-related genes may play different roles in the response of MSCs to mechanical stimulation.

Alkaline Phosphatase ; analysis ; Animals ; Antigens, Surface ; analysis ; Biomechanical Phenomena ; Bone Marrow Cells ; physiology ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Proliferation ; Core Binding Factor Alpha 1 Subunit ; analysis ; Fibroblast Growth Factor 2 ; analysis ; Insulin-Like Growth Factor II ; analysis ; Mesenchymal Stromal Cells ; physiology ; Osteoblasts ; physiology ; Osteogenesis, Distraction ; Pluripotent Stem Cells ; physiology ; Proto-Oncogene Protein c-ets-1 ; analysis ; Rats ; Stress, Mechanical ; Transforming Growth Factor beta ; analysis ; Up-Regulation ; physiology

ZM-1 tissue microarrayer designed by our group is manufactured in stainless steel and brass. It features an easier and faster preparation for tissue microarrays. By means of it, a group of biopsy needles are used to punch the donor tissue specimens respectively, and all the needles with the punched specimen cylinders are arranged into the array-board, where small holes have been digged to fit the needles. All the specimen cylinders arraying and the tissue microarray block's shaping are finished simultaneously. ZM-1 tissue microarrayer with a lower cost of manufacture, is capable of preparing the tissue microarrays conveniently, efficiently and quality-controllably.
Equipment Design ; Tissue Array Analysis ; instrumentation