Child ; Fibroma ; Humans ; Male ; Nasal Cavity ; pathology ; Nose Neoplasms

To study the drug-drug interaction of morinidazole and warfarin and its application, a sensitive and rapid liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of R-warfarin/S-warfarin in human plasma. In a random, two-period crossover study, 12 healthy volunteers received a single oral dose of 5 mg racemic warfarin in the absence and presence of morinidazole. Blood samples were collected according to a pre-designed time schedule. R-warfarin, S-warfarin and methyclothiazide were extracted with ethylether : methylenechloride (3 : 2), then separated on a Astec Chirobiotic V (150 mm x 4.6 mm ID, 5 microm) column using 5 mmol x L(-1) ammonium acetate (pH 4.0) - acetonitrile as mobile phase at a flow-rate of 1.5 mL x min(-1). The mobile phase was splitted and 0.5 mL x min(-1) was introduced into MS. A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the negative ion mode. Quantification was performed using multiple reaction monitoring (MRM). The resolution of warfarin enantiomers is 1.56. The linear calibration curves for R-warfarin and S-warfarin both were obtained in the concentration range of 5 - 1 000 ng x mL(-1). Intra- and inter-day relative standard deviation (RSD) for R-warfarin and S-warfarin over the entire concentration range across three validation runs was both less than 10%, and relative error (RE) ranged from -4.9% to 0.7%, separately. The method herein described is effective and convenient, and suitable for the study of metabolic interaction between morinidazole and warfarin. The results showed that coadministration of warfarin with morinidazole did not affect the pharmacokinetics of either R-warfarin or S-warfarin.

Objective To study the probability of transferring the regulatory T cells induced by TGF-β1 to prolong the allograft survival and the mechanisms involved.Methods According to the different culture conditions.three experimental groups were established:control group(T cells from C57 BL/6 mice cultured with II-2),MLR group(T cells from C57BL/6 mice activated by alloantigen)and TGF-βgroup(T cells from C57BL/6 mice activated by alloantigen and cultured with 5.0 ng/ml TGF-β1).After the culture,the ratio of CI4+CD25+T and the Foxq3 expression were measured by FACS and RT-PCR,respectively.On 9th day,the pathologic analysis was performed and the ratios of TH1,TH2 and Treg and the proliferation of lymphocytes were measured.Results The ratio of CD4+CD25+T in TGF-β group was higher than that in control group and MLR group(P<0.05),and Foxp3 was expressed in CD4+CD25+T cell from TGF-βgroup.After transferring ofthe cells,the allografi survival time in TGF-β group was prolonged and its mean survival time(MST)was(22.8±1.9)d,which was longer than that in MLR group and control group (P0.05),lower than that of control group(P<0.05). And the ratio of CD4+CD25+T in TGF-β group was higher than that in control and MLR group obviously(P

BACKGROUND: Recently, several scientists have found that differentiation of neural stem cells (NSCs) towards dopaminergic neurons may be increased in vitro by combination of some special cytokines. They have also found that dopaminergic neurons differentiated from NSCs can be used for the treatment of Parkinsn's disease. To improve the therapeutic effects of/n vitro transplantation, we should further study the biologic characteristics of NSCs at the induction and differentiation.OBJECTIVE: To explore the differentiation of NSCs which were incubated in differentiation solution for different time towards dopaminergic neurons in vitro.DESIGN: Single sample observation.SETTING: Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University.MATERIALS: This study was performed at the Department of Ncurosurgery, First Affiliated Hospital of Sun Yat-sen University from May to October 2007. Six healthy Sprague Dawley (SD) rats, gestational age 14 days, of clean grade, weighing 350-400 g,were provided by the Laboratory Animal Center, Sun Yat-sen University[permission No. SCXK (yue)2007-0034]. The protocol was performed in accordance with ethical guidelines for the use and care of animals.METHODS: NSCs derived from rat embryonic mesencephalon were cultured in serum-free culture medium containing epidermal growth factors and basic fibroblast growth factors. After passage, the NSCs were induced to differentiate towards dopaminergic neurons in the differentiation medium supplemented with interleukin 1o, interleukinl 1, human leukaemia inhibitory factors, and glial cell line-derived neurotrophic factors. The percentage of tyrosine hydroxylase positive neurons in differentiated ceils was detected with flow cytometer when NSCs were cultured in differentiation solution for 2, 4, 6, 8 and 10 days, respectively.MAIN OUTCOME MEASURES: Cellular morphological alteration of rat NSCs after differentiation. The percentage of tyrosine hydroxylase positive neurons in differentiated cells derived from NSCs.RESULTS: In differentiation medium, NSC spheres attached the bottom of plates and began to collapse. Cells inside the spheres grew out gradually and became irregular in shape. Six days later, most of the cells had I or 2 long processes and a few short processes. The percentage of tyrosine hydroxylase positive neurons in differentiated cells was respectively (3.2_+_0.9)%, (6.8 +1.6)%, (16.7-+2.6)%, (14.8_+1.8)% and (12.2_+2.5)% after culture for 2, 4, 6, 8 and 10 days, with significant differences (F =26.449, P ＜ 0.05).CONCLUSION: Induction time influences the differentiation of NSCs towards dopaminergic neurons in vitro. The percentage of dopaminergic neurons is the highest in differentiated cells derived from NSCs which are cultured in differentiation solution for 6 days.

BACKGROUND: Differentiation inducing factors and gestational age influence the differentiation potential of embryonic neuralstem cells.OBJECTIVE: This study was designed to observe the differentiation potential of rat mesencephalic neural stem cells at differentgestational ages towards dopaminergic neurons.DESIGN: A randomized controlled observation.SETTING: Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou City, GuangdongProvince, China.MATERIALS: This study was performed at the Laboratory of the First Affiliated Hospital of Sun Yat-sen University between Marchand September 2007. Thirty adult gestational SD rats, weighing 350 400 g, were provided by the Laboratory Animal Center of SunYat-sen University (Permission No. 2007-0034). The protocol was performed in accordance with ethical guidelines stated in Guide forthe use and care of laboratory animals, approved by the Committee on the Care and Use of Laboratory Animals of the Institute ofLaboratory Animal Resources Commission on Life Sciences, National Research Council, China (1985). DMEM/F12 serum-free medium,B27 additives, epidermal growth factor, basic fibmblast growth factor, and fetal bovine serum (volume fraction:0, 1) were purchased fromGibco Company, British; Interleukin lα, interleukin 11, and glial cell-derived neurotrophic factors were purchased from R&D Company,USA; In addition, leukaemia inhibitory factor (Perpotech, British), tyrosine hydroxylase(Santa Cruz, USA), nidogen antibody,microtubule-associated protein 2 antibody, and glial fibrillary acidic protein antibody(Chemicon, USA) were also used.METHODS: Six rats were randomly selected at each time point (on days 10,12,14,16, and 18 after gestation). After anesthesia, therats were sacrificed. Under the aseptic condition, fetal rat was harvested. Rat mesencephalic ventral brain tissue was isolated forculture of neural stem cells. Different gestational ages of rat brain-derived neural stem cells were separately cultured in theserum-free medium containing epidermal growth factors and basic fibroblast growth factors. After passage and amplification, theneural stem cells were induced to differentiate towards dopaminergic neurons in the medium containing interleukin lu, interleukin11, leukaemia inhibitory factors, glial cell-derived leukaemia inhibitory factors. On day 6 after induction and differentiation, thedopaminergic neurons were observed and identified by immunocytochemistry. After labeled by tyrosine hydroxylase, thedifferentiated dopaminergic neuron proportion was detected by a flow cytometer.MAIN OUTCOME MEASURES: The growth state of differentiated rat neural stem cells at different gestational ages and theimmunocytochemistry results. The tyrosine hydroxylase staining-positive neural stem cell proportion after induction anddifferentiation.RESULTS: Rat mesencephalic neural stem cell spheres on days 10,12, 14, 16, and 18 after gestation adhesively grew in thedifferentiation-inducing medium. The neural stem cells in the spheres gradually grew in radial tendency. On day 6 afterdifferentiation, most of the neural stem cells exhibited 1-2 long processes or several short processes. After nidogenimmunocytochemical staining, most of neural stem cells exhibited cytoplasm-positive. After culture for 6 days in the differentiationinducing medium, rat mesencephalic neural stem cells at gestational 10,12, 14, 16, and 18 days were detected by a flow cytometer.Results demonstrated that the proportion of tyrosine hydroxylase-positive cells was (10.3±2.5)%, (21.6±3.4)%, (16.7±2.8)%,(14.2±3.2)%, and (8.9±1.8)%, respectively. There was a significant difference in the proportion of tyrosine hydroxylase-positivecells among the cells at different gestational days (P < 0.05). Rat neural stem cells at gestational 12 days could be induced todifferentiate into dopaminergic neurons at the highest proportion.CONCLUSION: Mesencephalic neural stem cells of rats at different gestational days have different capabilities to differentiatetowards dopaminergic neurons. The proportion of dopaminergic neurons is the highest when mesencephalic neural stem cells ofrats at gestational 12 days.

BACKGROUND: It is a key point to revascularize the tissue-engineered bone during the repairing of large bone defect. Fascia flap is commonly used in clinic to accelerate the blood supply of implant.OBJECTIVE: To observe the feasibility of repairing goat tibia defects with tissue-engineered bone and accelerating revascularization with fascia flaps.DESIGN: Randomized and controlled animal experiment SETTING: Department of Traumatic Orthopaedics, Nanfang Hospital,Southern Medical University.MATERIALS: Totally 36 goats with the body mass of 14.5-15.5 kg of either gender were enrolled.METHODS: This experiment was conducted at the Department of Traumatic Orthopaedics, Nanfang Hospital, formerly the First Military Medical University of Chinese PLA from December 1999 and December 2003.Bone and periosteum defects 20 mm long were made and fixed with plate of left tibia in 36 goats. They were randomly divided into four groups: Group A in which the defects were filled with coral hydroxyapatite (CHAP), Group B I CHAP+ bone marrow stroma cells (BMSc); Group C with fascia flaps;Group D with nothing. Next, the bone regeneration and the revasculariza tion were evaluated. Radionuclide bone imaging was done 2, 4, 8 weeks after operation. After X-ray examination, the index of optical density of Xray films and histology of the implants were analyzed at 4, 8, 12 weeks after operation, and the biomechanical characters were studied 12 weeks postoperatively.MAIN OUTCOME MEASURES: Gross observation and X-ray, radionuclide bone imaging, biomechanical and histological observation RESULTS: Totally 36 goats entered the stage of result analysis. ① Gross observation of the repair sample of bone defects of the animals in each group: there was no osteogenesis postoperatively at each time point in the blank control group . In Group B, at week 8 to 12, there was no obvious osteogenesis and callus formation on the surface of the materials. In Group C,At weeks 8 to 12, bone defects were filled gradually, many bone callus processes were seen on the surface of the materials , centralizing and enwrapping the materials. The osteogenetic process in the Group C were superior to that of theGroup B. ②Examination result with -901/SA PET-CT scanners: It was seen by naked eyes that at weeks 2 to 8 in the Group A,the radioactivity concentration at region of interesting (ROI) of the operation side had obvious increasing trend, and similar trend of changing appeared in the Group B and Group C, but the ROI counts and T/NT value in the Group B were both lower than those in the Group C. The decreasing trend in the Group A was lower than that in the Group B. ③) Radiological results: the osteogenesis volume through measuring absorbance in the order from large to small was Group C, Group B, and Group A[At week 12, they were (4.180±0.192), (3.480±0.453), (2.959±0.682)respectively ].④Biomechanical results: there were significant difference of loading and bending stress in the Group C, Group B and Group A [ The loading was (758.333±88.754), (530.214±65.297), (359.667±60.715)N , respectively; and the bending stress was (13.937±2.199), (10.123±1.243),(6.223±0.945)N/mm2, respectively ].⑤)Histological results: Slices at various time points in the blank control group showed no bone tissue. In the other three 3 groups, with the prolongation of time, the osteagenetic range and quality were in the order of Group C, Group B and Group A.CONCLUSION: The fascia flaps can accelerate the revascularization process in the formation of tissue-engineered bone so that the capability of tissue engineered bone to repair the large bone defects may be enhanced.

BACKGROUND: The main aspect of the study in the bone histological engineering is how to maintain and improve theosteogenesis of the osteoblasts in vivo and in vitro. The gene transference may provide a new effective method to deal with theproblem.OBJECTIVE: To discuss the effect of the reverse transcription virus mediated human bone morphogenetic protein7(hBMP-7) gene transfection on the proliferation and osteogenetic differentiation of the bone marrow mesenehymal stemcells (BMSCs) of the rabbits.DESIGN:Cells taken as the study object, grouping control, repeat observation andmeasurement.SETTING: Traumatological and othopaedic lab of a medical university hospital.PARTICIPANTS: The study wascompleted in the Traumatological and Othopaedic Lab in the Affiliated Nanfang Hospital of the Southern Medical University from July 2001 to July 2003. Four New Zealand rabbits,whose weights varied from 1.0 to 1.5 kg, were provided without sexlimit by the Animal Experiment Center of the First Military Medical University of Chinese PLA.METHODS:The reverse transcription virus carriersof the hBMP-7 were constructed,and then the BMSCs were transfected by the virus containing target genes. The expression of the hBMP-7 protein was detected with the immunohistochemical method. The cell proliferation, cycle and ALP synthesis were respectively detected with the MTT method,flow cytometer and NPP method.MAIN OUTCOME MEASURES: Primary results: ① the detection results of the cell proliferation. ② the detection results of the ALP.Secondary results: ① the expression of the hBMP-7 protein in the transfected BMSCs. ② the detection results of the cell cycle.RESULTS: After the BMP-7 gene transfection, there was hBMP-7 positive expression in the BMSCs of the rabbits,using the immunohistochemical detection. There was no significant change in the BMSCs proliferation of the rabbits after the hBMP-7 gene transfection ( P ＞ 0.05). Compared with the ALP synthesis of the transfected BMSCs(294. 592 ± 86. 567) nkat/L, there was significant difference in the ALP synthesis of the empty carrier transfected BMSCs(155. 231 ±86.567) nkat/L and the un-transfected BMSCs (160. 866 ±91. 585)nkat/L( F =5. 660, P ＜ 0. 05).CONCLUSION: After the BMP-7 gene transfection, the BMSCs can synthesize and express the extragenous BMP-7. The hBMP gene transfection can promote the differentiation of the BMSCs cultured in vitro into the osteoblasts and can be used as the seed cells in the construction of the histological en gineering bone tissues and in further application.

Purpose Brain complications severely threaten the treatment and survival of children with leukemia. This paper aims to investigate the MRI manifestations and differences of brain complications in leukemia before and after chemotherapy for a clinical guidance.Materials and Methods The clinical data and MRI findings of 37 children with leukemia and brain complications were retrospectively analyzed. Thirty-four of them underwent MRI scan twice or more, among whom 28 received contrast-enhanced MRI scan.Results Twenty-two patients were discovered with brain complications before chemotherapy, 2 of whom were with two kinds of complications. Meningopathy was found in 7 patients who showed widespread or localized meningeal thickening. Among them, 5 patients'' lesions reduced or disappeared after chemotherapy. Intracerebral multiple small and micro bleed was found in these 7 patients, 2 of them combined with hematoma. Three patients were found with intracranial tumor which all proved to be temporal bone tumor, 1 of whom combined with temporal lobe tumor and 1 had tumor disappeared after chemotherapy. The other complications before chemotherapy included leukoencephalopathy (n=2), subdural collection of fluid (n=2), meninges and parenchymal infiltration of leukemia (n=1), fungal infection (n=1) and cerebral infarction (n=1). On the contrary, 17 patients were discovered with brain complications after chemotherapy, 8 of whom were with two or more complications. Two patients had different kinds of complications before and after chemotherapy. Brain atrophy was observed in 13 patients. Leukoenphalopathy was found in 9 patients who presented high signal in white matter of double periventricular and/or semi-oval center on T2WI; the lesions of 4 patients were reduced or disappeared after withdrawal. Infectious diseases were diagnosed in 3 patients, including viral encephalitis in 2 cases, tuberculous meningitis combined with tuberculoma in 1 case. The other complications included intracranial tumor (n=2), sinus thrombosis (n=1), posterior reversible encephalopathy syndrome (n=1) after chemotherapy. Conclusion The MRI findings of brain complications in childhood leukemia are various and demonstrate significantly different features before and after chemotherapy. The major complications before treatment include meningopathy and intra-cerebral hemorrhage;while after chemotherapy the main complications are brain atrophy, leukoencephalopathy and infectious diseases. MRI proves to be a valuable method to detect, observe and follow up these complications.

Objective To observe the clinical effects of the adjustive tractor for cervical dislocation.Methods Forty-seven patients were included between September 2007 and November 2011.There were 36 males and 11 females with age ranged from 7 to 62 (mean,35 years).The mean interval from injury to admission was 8.6 h (range,0.5-72 h).There were atlanto-occipital dislocation in 2 cases,C1.2 in 2 cases,C2.3 dislocation in 5 cases,C3.4 dislocation in 2,C4.5 in 9,C5.6 in 13,C6.7 in 14.Thirty cases were complicated by fracture.No facet locking occurred in sixteen cases.Facet locking was found in ten cases and bilateral facet locking was in 21 cases.After reduction,brace or internal fixation followed.According to American Spinal Injury Association (ASIA) spinal function impairment scale standard,there were 4 cases in level A,10 cases in level B,18 in C,10 in D,and 5 in E.According to Japanese Orthopaedic Association (JOA) spinal function rating standard,the mean JOA score was 9 (range,2-14).Results All 47 cases were reduced successfully without neuronal function aggravation.Traction power ranged from 7 to 60 kg (mean,25.6 kg),the mean time of traction was 8 min (range,3-10 min).The mean follow-up was 38 (range,6-48) months.All the patients achieved normal alignment and intervertebral height.The intervertebral body fusion was observed in all of cases,the mean fusion time was 3.3 months (range,3-6 months).One patient who experienced nonunion of vertebral arch fracture didn't receive further treatment because of absence of symptoms.At last follow-up,there were 3 cases in level A,1 in level B,4 in C,8 in D,and 31 in E according to ASIA scale.The mean JOA score was 12 (range,2-17).Conclusion The adjustive tractor is simple and safe for prompt reduction.

OBJECTIVETo observe the effects of Astragalus and Angelica on bone marrow stem cells (BMSU) proliteratlon mn vitro and investigate its possible mechanism.

METHODSFive 200 to 220 g SD rats were fed with a high fat diet for 4 weeks and given 30 mg/kg streptozotocin (STZ) twice develop type II diabetes from July 2009 to February 2010. The rats with blood glucose concentrations of 16.7 mmol/L or more were considered diabetic. Bone Marrow Stem Cells (BMSC) were collected and isolated by density gradient centrifugation. The BMSC were divided into 4 groups,including empty control group, Astragalus group, Angelica group and Astragalus plus Angelica group. DMEM of 100 microl was added in empty control group. DMEM of 100 microl containing Astragalus (1100 mg/L), Angelica (1100 mg/L) and Astragalus (1100 mg/L) combine with Angelica(220 mg/L) were added in Astragalus group, Angelica group and Astragalus plus Angelica group respectively. The cell proliferation was detected by MTT method, and the concentration of VEGF in the supernatant was determined by ELISA. The VEGF expression was analyzed by Western Blot after 14 days incubation.

RESULTSThe BMSC proliferation and the VEGF concentration in the supernatant and the BMSC VEGF protein expression significantly increased in Astragalus group and Astragalus plus Angelica group compared to those of empty control group (P < 0.05 or P < 0.01). The above effects were more strong in Astragalus plus Angelica group (P < 0.05).

CONCLUSIONAstragalus with Angelica or used separately could promote BMSC proliferation. The mechanism might induce the VEGF protein expression in BMSC. And the independent use of Angelica has no above effect.

Angelica ; Animals ; Astragalus Plant ; Bone Marrow Cells ; cytology ; drug effects ; Cell Proliferation ; drug effects ; Diabetes Mellitus, Experimental ; drug therapy ; Hematopoietic Stem Cells ; cytology ; drug effects ; Male ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; analysis