Diagnosis, Differential ; Female ; Humans ; Infant, Newborn ; Nasopharyngeal Neoplasms ; pathology ; surgery ; Polyps ; pathology ; surgery ; Teratoma ; pathology

Dibenzothiophene (DBT) was used as a model compound. A bacteria strain, which can degrade dibenzo-thiophene efficiently, was obtained. This strain was identified as Pseudomonas stutzeris UP-1 according to its morphological, physiological and biochemical characters, and 16S rDNA sequence. The strain exhibits strong degradation capacity of DBT, and the end product of degradation is a kind of soluble compound. After the analysis of product of DBT degradation, it was deduced that the degradation of DBT by Pseudomonas stutzeri UP-1 is in accordance with the Kodama mechanism.

AIM: To investigate the potassium channel gene expression of myocardial sleeves of pulmonary vein and effects of amiodarone on rabbits with rapid atrial pacing.METHODS: Rabbits were divided into three groups(n=10),(1) the control group with sham operation and placebo;(2) the right atrial pacing(RAP) group at 600 beats/min with the placebo;(3) the amiodarone group treated for seven days with oral amiodarone at 100 mg ? kg-1 ? d-1.Based on RAP simultaneously,the messenger ribonucleic acid(mRNA) of specimen was measured by reverse transcription-polymerase chain reaction.RESULTS: Compared with the control group,Kv4.3(transient outward K+ current,Ito1) mRNA expression in RAP group was reduced by 51%(P

AIM: To study the effect of vascular endothelial growth factor(VEGF) on hematopoietic differentiation from mouse embryonic stem cells(ESC) in vitro.METHODS: ES-D3 was allowed to grow on mouse fetal fibroblast feeder layer,and then was developed into embryoid bodies(EB).EB cells were transferred into medium supplemented with different concentration of VEGF and VEGF+SCF for 1 week.Six groups,including.VEGF 5 ?g/L,VEGF 10 ?g/L,VEGF 20 ?g/L, VEGF 5 ?g/L+SCF,VEGF 10 ?g/L+SCF and VEGF 20 ?g/L+SCF,were designed.The group of spontaneous differentiation without cytokine(s) was used as control.Hematopoietic transcription factor GATA-2 and early hematopoietic differentiation genes(c-kit and ?-H1) were detected by RT-PCR.The content of CD34~+ cells in each group were measured by flow cytometry.The cells derived from ESC were incubated in semisolid methycellulose cultures.The numbers of total colony-forming units in culture(CFU-C) were counted by reverse microscope.RESULTS: ES-D3 grew and formed EB at day 4.VEGF had a stimulatory effect as a single factor on the expression of genes associated with early hematopoietic differentiation(GATA-2,c-kit and ?-H1),the generation of CD34~+ cells and CFU-C in EB.The effects of VEGF+SCF were the most potent in the experimental groups according to the percent of CD34~+ cells and the numbers of hematopoietic colonies.The most highest inducing efficacy was achieved in VEGF 20 ?g/L or VEGF 10 ?g/L combined with SCF.CONCLUSION: VEGF promotes the differentiation of ESC into hematopoietic cells in vitro.The strongest effect was achieved when VEGF was combined with SCF.

To produce antibodies capable of neutralizing botulinum neurotoxin type B(BoNT/B),We cloned the carboxy-terminal end of Hc containing the major determinants responsible for specific toxin,induced and purifed.The heavy-chain and kappa light-chain variable region gene repertoire of immunoglobulin were amplified individually from the spleen cell mRNA by RT-PCR and joined as a single-chain Fv(scFv)DNA fragment.These fragment were cloned into the phagemid pCANTAB5E and the phage display library was constructed.Results showed that the high affinity scFv was obtained after 4 rounds of panning,with its DNA sequence conforming to that of mouse antibody.

OBJECTIVETo explore the effect of arteriosclerosis obliterans (ASO) blood stasis syndrome (BSS) serum on vascular endothelial cell injury and to study the regulation of Taohong Siwu Decoction (TSD) on it.

METHODSUmbilical vein endothelial cell culture system was established. The serum endothelial cell injury model with ASO BSS was prepared. Low, medium, and high concentrations TSD containing serums were respectively added. The endothelial cell proliferation activity was observed by MTT method. Ultrastructures of endothelial cells were observed under transmission electron microscope. Changes of intracellular calcium ion concentration and the cytoskeleton were observed under laser confocal microscope. Contents of ET, NO, and transforming growth factor beta1 (TGF-beta1) in endothelial cell culture supernatant were detected by ELISA.

RESULTSIn ASO BSS serum group endothelial cell proliferation activities decreased, the cell structure was obviously destroyed, calcium ion concentration increased, contents of ET, NO and TGF-beta1 increased significantly (P < 0.01), and ET/NO ratio was imbalanced. After incubating with TSD drug containing serum, endothelial cell proliferation activities and injured cell structures were obviously improved; ET, NO and TGF-beta1 levels decreased (P < 0.05, P < 0.01), ET/NO ratios approximated to the normal level.

CONCLUSIONThe main mechanism of TSD for treating ASO ASS lied in improving injured vascular endothelial cells and endocrine disorder.

Arteriosclerosis Obliterans ; Cell Proliferation ; Drugs, Chinese Herbal ; therapeutic use ; Endothelial Cells ; Humans ; Medicine, Chinese Traditional ; Serum ; Transforming Growth Factor beta1 ; metabolism ; Umbilical Veins

OBJECTIVECord blood (CB) provides a rich source of stem cells for transplantation. CB transplantation has been used widely after myeloablative therapy. One major disadvantage of CB transplantation is delayed platelet engraftment. The aim of this study was to hasten platelet engraftment by investigating the ability of different hematopoietic growth factor combinations to generate large numbers of megakaryocyte (Mk) from CB and by evaluating the biologic characteristics and function of the expanded Mk.

METHODSCB samples were obtained at the end of normal full-term deliveries with informed consent. Mononuclear cells (MNCs) were isolated from CB using Ficoll density centrifugation. MNC population was positively selected for CD(34) expression by magnetic cell sorting (MACS). CD(34)(+) cells were cultured in serum-free and stroma-free medium containing the following two different cytokine combinations: thrombopoietin (TPO) + stem cell factor (SCF) + interleukin (IL) -3 + IL-6 and TPO + SCF. Cultures were characterized after 3, 7, 10 and 14 days by flow cytometry, colony forming unit-megakaryocyte (CFU-Mk) and maturation evaluation (Mk ploidy). The expanded Mk function was examined by the platelet activation in vitro and severe combined immunodiffiency (SCID) mice transplantation in vivo.

RESULTSDifferent results were observed with different culture conditions. With the first cytokine combination optimal expansion of CD(41)(+) cells was observed on day 10, but the optimal expansion of Mk progenitors (CD(34)(+)CD(41)(+)) was observed on day 7, with a median 121 and 44-fold increase at the starting cell dose. This result was also proven by CFU-Mk. The largest numbers of CFU-Mk were also observed on day 7. The degree of maturation of Mk cells also increased as suggested by DNA content of CD(41)(+) cells, which means that CD(34)(+) cells cultured for 3 - 7 days were richer in primitive Mks, while those cultured for 10 - 14 days had greater numbers of more differentiated Mks. For the second cytokine combination, CD(41)(+) and CD(34)(+)CD(41)(+) cells were fewer than the first one, but it produced 36 and 85-fold CD(34)(+)CD(41)(+) and CD(41)(+) respectively on day 7. Platelet activation test confirmed that the expanded Mks had normal function. Therefore, the expanded Mks could be transplanted into the SCID mice bone marrow and produce human platelet in the peripheral blood of the mice.

CONCLUSIONEx vivo expanded Mk might facilitate CB transplantation and help shorten the period of post-transplant thrombocytopenia.

Animals ; Antigens, CD34 ; Cell Culture Techniques ; methods ; Cells, Cultured ; Culture Media ; Fetal Blood ; cytology ; Humans ; Leukocytes, Mononuclear ; cytology ; Megakaryocytes ; cytology ; Mice ; Mice, SCID

OBJECTIVETo develop a procedure by which Schwann cells and myelin in the peripheral nerve could be removed while the basal lamina tubes remained intact, and to obtain a thick and long acellular nerve allograft in humans.

METHODSFour ulnar nerves 10.0 cm long and 4.0 - 5.0 mm in diameter were excised from a donated male body and cleaned from external debris. The nerves were treated with a solution of Triton X-100 and a solution of sodium deoxycholate at room temperature. After a final wash in water, the nerves were stored in phosphate-buffered saline (PBS, pH 7.2) at 4 degrees C. HE, luxol fast blue and fibrin staining were performed to visualize cells, myelin and basal membranes respectively and immunohistochemical staining was performed to visualize the presence of laminin, a Schwann cell lamina component, both in fresh and acellular nerve segments. To reveal overall structure better, methylene blue-fuchsin staining was performed in semithin section. The ultrastructure of acellular and fresh nerves were observed and photographed in a transmission electron microscope.

RESULTSThe acellular human ulnar nerve was white long cylinder with well elasticity and ductility. HE, myelin and fibrin staining revealed that cells, axons and myelin sheath were removed and basal membrane was preserved after extraction procedure. Staining for the presence of laminin showed that the Schwann cell basal lamina component were present in the nerves after chemical treatment. Methylene blue-fuchsin staining and transmission electron microscopy showed that the myelin sheaths were absent in the extracted nerve segments and empty basal lamina tubes remained in the endoneurium.

CONCLUSIONSWe developed an extracted procedure with the detergents of Triton X-100 and deoxycholate, by which cells, axons and myelin sheaths could be removed from a human ulnar nerve while the basal lamina tubes remain intact and a thick long acellular nerve allograft is obtained. The laminin, a Schwann cell basal lamina component, can be preserved in the acellular nerve.

Adult ; Axons ; drug effects ; Cell Separation ; methods ; Deoxycholic Acid ; pharmacology ; Humans ; Male ; Myelin Sheath ; drug effects ; Octoxynol ; pharmacology ; Transplantation, Homologous ; Ulnar Nerve ; cytology ; transplantation ; ultrastructure

OBJECTIVETo evaluate the feasibility and characteristics of human engraftment in HLA disparate cord blood transplantation.

METHODSTwo human HLA-haploidentical or HLA-mismatched cord blood units were transplanted into sublethally irradiated severe combined immunodeficiency (SCID) mice. The characteristics of engraftment, hematopoietic and immunological reconstitution between the two groups were compared.

RESULTSTwo mixed cord blood units can engraft in SCID mice with donor-recipient chimerism and reconstitute hematopoiesis and immunological functions. No unfavorable factors had been observed. Only one of the two cord blood units which had higher colony forming ability in vitro could engraft in most SCID mice as shown by HLA-DQB(1) gene detection. Two HLA-haploidentical cord blood units were simultaneously engrafted in 3 SCID mice.

CONCLUSIONDouble HLA-haploidentical or HLA-mismatched cord blood can engraft in SCID mice and reconstitute hematopoietic and immunological functions. HLA disparity has no significant effect on survival and engrafting rate. However, in less HLA disparity group, two cord blood units were prone to engraft simultaneously.

Animals ; Antigens, CD ; immunology ; Cord Blood Stem Cell Transplantation ; methods ; Disease Models, Animal ; Female ; Fetal Blood ; immunology ; metabolism ; Flow Cytometry ; HLA Antigens ; genetics ; immunology ; Hematopoiesis ; Humans ; Mice ; Mice, SCID ; Random Allocation ; Severe Combined Immunodeficiency ; immunology ; physiopathology ; surgery ; Survival Analysis ; Transplantation, Heterologous

OBJECTIVETo investigate the feasibility of hip arthroplasty in the treatment of elderly patients with Evans I-III intertrochanteric fracture of femur by analyzing its biomechanics characters.

METHODSWe solved the CT digital image files with the graphics processing software Mimics at DICOM 3.0 standard, and reconstructed the three-dimensional entity of femur with CAD modeling software Unigraphics. Then the fracture line was defined in the model as the line between the tip of greater trochanter and inferior margin of small trochanter, above which the upper bone was removed. Afterwards the two prosthesises with different stem lengths (120 mm and 170 mm) were implanted into the fracture model respectively as hip arthroplasty with 3 mm bone cement layer between prosthesis and femur, and the bone defect was repatched with 5 mm bone cement layer. A three-dimensional finite element model was established with finite element analysis software ABAQUS 6.5. We formulated different material parameters under the stress condition standing with single leg to build the stress distribution map of the femur prosthesis, and took 5 loci of region of stress concentration to calculate the mean value of stress.

RESULTSThe stress distribution maps of the short and long stem length prothesises were similar. And there were two areas of stress concentration, including the upper portion and the lower portion close to the joint of the prosthesis stem, and the stress concentration in the junction part was obviously between the lower portion and the upper area of the small trachanter. The stress reached the first concentration area at the junction and then gradually reached the second concentration area at the interior terminal of the stem. While the stress gradually increased along the lateral prosthesis stem, and reached the stress concentration area at the end.

CONCLUSIONSThe stress distribution maps in the femur prosthesises are similar between hip arthroplasty in the treatment of intertrochanteric fracture of femur and the traditional hip arthroplasty surgery. The peak stress values are higher in the long stem prosthesis in the treatment of intertrochanteric fracture of femur than the short type, while they are under the rupture value of the metal.

Aged ; Arthroplasty, Replacement, Hip ; instrumentation ; methods ; Biomechanical Phenomena ; Bone Cements ; Computer Simulation ; Female ; Finite Element Analysis ; Hip Fractures ; surgery ; Hip Prosthesis ; Humans ; Image Processing, Computer-Assisted ; Software ; Stress, Mechanical