The quantitative parameters based on dynamic contrast-enhanced MRI (DCE-MRI)scan,simulate the distribution of contrast inside and outside of the blood vessels through a variety of tracer kinetic models.The DCE-MRI perfusion parameters now are widely applied in clinical management of uterine tumors,to analyze the microcirculation characteristics in tumor and to guide the diagnosis,tumor grading and efficacy evaluation.

Objective To assess dermatoscopic characteristics of lichen sclerosus(LS). Methods Dermatoscopy was performed to observe 27 genital or extragenital porcelain-white skin lesions in 15 outpatients with confirmed LS. Results As dermatoscopy showed, of the 27 skin lesions, 24 showed white-yellowish structureless areas, 25 linear vessels, 27 white patches, 17 keratotic plugs. Characteristic cloverleaf-like structure was observed in 7 skin lesions in 4 patients, which was consistent with LS. Conclusion Patients with LS show atypical clinical manifestations, but specific dermatoscopic patterns.

Objective To provide methodological references for laboratories to carry out newborn screening for disorders of amino acid metabolism,we compared the difference and distribution of ten amino acids including alanine (Ala),arginine (Arg),citrulline (Cit),glycine (Gly),leucine (Leu),methionine (Met),ornithine (Orn),phenylalanine (Phe),tyrosine (Tyr),and valine (Val) from newborn dried blood spots specimen using derivatization or non-derivatization as sample preparation methods.Method It is a comparative research study.A total of 4135 newborn screening dried blood spot samples for inborn errors of metabolism were collected from January to June,2012.All specimens came from neonatal screening center of shanghai children's hospital.Samples were prepared by two different techniques,the corresponding kits and the procedures were used as follows:(1) Simultaneous detection of 100 dried blood spot specimens using two methods respectively to compare the paired difference of each amino acid.(2) 2000 cases of normal newborn specimens were detected respectively to obtain the normal distribution of ten neonatal amino acids.(3) 35 specimens from patients previously diagnosed positively as inborn errors of metabolism were simultaneously detected with 7 amino acids to verify the consistency of two techniques in clinical judgment.Results The amino acid levels of normal newborns analyzed by one-sample.kolmogorovSmirnov test (Z value ranged from 1.997 to 6.229) showed a skewed distribution (P ＜ 0.01).Except for Leu and Tyr,non-derivatization techniqueshowed a lower concentration than derivatization technique,and the CVs of nine amino acids were ＜ 10％ except for Met (the CV of Met was 47.8％),and the average CV is 7.8％.Except for Met,Phe and Tyr,the levels of other 7 amino acids measured by two techniques showed no significant difference (P ＞ 0.05).According to 0.5th to 99.5th percentiles,the normal reference range for derivatization method were greater than on-derivatization method,and the average value was 25.3％.After clinic judgment,the results of the abnormal indicators of children with true metabolism disorders showed no statistically significant between two methods (P ＞ 0.05),the detection rate was 100％.Conclusions There was a slight difference between derivatization and the non-derivatization techniques in detecting multiple amino acids.The results of the abnormal indicators of amino acid metabolism disordersshow no statically significant difference between the two methods,and no difference in clinical judgment.Both methods can be used in detecting amino acid metabolism disorders in newborn screening.

Objective To summarize the acylcarnitine profile in children with malnutrition,with an attempt to distinguish it from those of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency,multiple acylCoA dehydrogenase (MAD) deficiency,or glutaric aciduria type Ⅱ (GA Ⅱ).Methods Thirteen pediatric patients with malnutrition and 214 children of the same age but without malnutrition,which was set as the control group,were included in this study.The blood samples were collected at admission,and the concentration of carnitine and acylcarnitines were measured in bloodspots by tandem mass spectrometry using samples nnderivatized.Results The concentrations of acylcarnitines which were involved in fatty acid oxidation,including octadecanoyl (C18) to acetyl (C2) acylcarnitines and ketonic acylcarnitines,were higher in malnutrition group than in the control group.Particularly,the concentration of decanoyl acylcarnitine (C10) in the malnutrition group was (0.203 ±0.105) μmol/L,which was out of the normal rang (0-0.200 μmol/L),was significantly higher than that [(0.054 ±0.030) μmol/L] in the control group (P ＜0.001).There was no significant difference in the concentrations of acylcarnitines [e.g.propionyl (C3),isovaleryl (C5),3-hydroxy-isovaleryl (C5OH),and glutaryl (C5DC) acylcamitines] involved in amino acid decomposition between the malnutrition and control groups.Conclusions The concentrations of acylcarnitines related to fatty acid oxidation elevate in children with malnutrition.In particular,the medium-chain acylcarnitines C10 is out of the normal range,which can be used to differentiate malnutrition from MCAD and MAD.

BACKGROUND: How to reconstruct tissue-engineered bone with structure similar to natural bone iS a problem in the development of tissue engineering. Cell sheet engineering technology enables novel approaches to construction of tissue-engineered bone. OBJECTIVE: To observe the biocompatibility of call sheets to decalcified bone matrix (DBM) and their growth on DBM. DESIGN, TIME AND SETTING: An in vitro observation was performed at the Central Laboratory, Affiliated Hospital, Qingdao University Medical College between June and September 2009.MATERIALS: Dog bone marrow stromal cell sheets were prepared using temperatura-responsive medium. Dog DBM was prepared by defatting, decalcification, and noncotlagen protein removal procedures. METHODS: DBM surface was covered by call sheets prepared by temperature-responsive technology and cultured with DMEM containing 10% fetal bovine serum and osteoinductive agent.MAIN OUTCOME MEASURES: Under scanning electron microscope, DBM structure, as well as the attachment and growth of cell sheets on DBM surface, was observed. Porosity and aperture size of DBM were calculated. RESULTS: DBM exhibited a three-dimensional latticed structure, with a porosity of approximately 75%. The mean aperture size was (250.11±98.89) μm, exhibiting a normal distribution. Cell sheets well attached to and grew on DBM surface, and rapidly proliferated.CONCLUSION: Cell sheets show good biocompatibility to DBM. DBM/cell sheets complex can be applied in tissue-engineered bones, which promotes the construction of tissue-engineered bone with structure similar to natural bone.

Objective: To confirm the structure and preferential conformation of the Schiff base of gossypol with 1, 3, 4, 6-tetra-O-acetyl-β-D-glucosamine and explore its anti-HIV-1 activity.Methods: The Schiff base of gossypol with 11, 3, 4, 6-tetra-O-acetyl-β-D-glucosamine was synthesized and identified by FT-IR, NMR spectroscopy and the PM6 semi-classical calculation.The inhibitory activity of the novel compound against the laboratory-adapted HIV-1IIIB strain was examined using the HIV-1IIIB/TZM-bl indicator cell culture system.Results: The 1H and 13C-NMR signals of the new Schiff base were assigned.The PM6 semi-classical calculation indicated that enamine-enamine tautomeric form of the new Schiff base was more stable,which was stabilized by the intramolecular hydrogen bonds.The anti-HIV-1 test showed that the compound could block the entry of HIV-1IIIB into the target cells.Conclusion: The Schiff base of gossypol with 1, 3, 4, 6-tetra-O-acetyl-β-D-glucosamine exhibits enamine-enamine tautomeric form in solution, which shows potential anti-HIV-1 activity.

Objective Diffusion/perfusion-weighted MRI (DWI、PWI) was performed to evaluate growth and vascularity of implanted C6 rat gliomas. Methods 36 female Wistar rats were implanted of C6 glioma cells intracerebrally. Between 1 and 4 weeks after implantation, eight to ten different rats were imaged with T_1WI, T_2Weighted imaging, DWI, PWI, and postcontrast T_1WI at each weekly time point. After MRI, each rat brain was examined histologically using HE and immunohistochemical staining for CD34. Results On DWIs, statistical differences of apparent diffusion coefficient (ADC) values for both the solid tumor component and peritumoral region were present comparing 3-4 weeks with 1-2 weeks after implantation (P

BACKGROUND:Conventional methods,including trypsin digestion and cells transfer using single call suspension,have many drawbacks,which limit the development of bone tissue engineering.OBJECTIVE:To culture bone marrow stromal stem calls,induce osteoblastic differentiation,and prepare cell sheets.METHODS:Canine bone marrow stromal calls were isolated by density gradient centrifugation technique,inoculated into DMEM medium,and induced to differentiate into osteoblasts.Complete call sheets were harvested by call sheet engineering based on the temperature change of temperature-responsive medium.RESULTS AND CONCLUSION:Immediately after inoculation,primary calls were scattered on the bottom of culture flask,presenting a transparent spherical body with a good refractive capacity.At 12 hours,calls exhibited a long shuttle shape,reached complete confluency,and grew in a whirlpool-like fashion.After osteoblastic induction,the majority of bone marrow stromal stem calls appeared tetragonal,polygonal,and squamose.At 21-28 days,round or oval-shaped calcified nodules formed.When the bone marrow stromal stem calls in the temperature-responsive culture dishes were cooled below the critical temperature 32℃,cells were gradually detached from the bottom of culture flask and formed complete bone marrow stromal stem call sheets.These findings indicate that density gradient cantrifugation technique can be used to successfully isolate and culture canine bone marrow stromal stem cells to differentiate into osteoblasts and call sheet engineering enables to harvest complete bone marrow stromal stem call sheets.

BACKGROUND:There are some disadvantages in harvesting and transferring cells in the traditional tissue engineering technique,and it is difficult to form dense tissues,which significantly limits the development of tissue engineering.OBJECTIVE:To explore the culture and fabrication of dog bone marrow mesenchymal stem cell(BMSC)sheet in vitro.METHODS:Bone marrow was extracted from dogs following anesthesia.BMSCs were isolated with the method of density gradient centrifugation in vitro.BMSCs at passage 4 at a density of 1×10~9/L were incubated in the temperature-responsive culture dishes with a diameter of 3.5 cm,and cultured in an incubator at 37 ℃,5% CO_2 and saturated humidity.The temperature of the incubator was changed from to 37 V to 20 ℃ to prepare BMSCs cell sheet for 20 minutes.Cell morphological changes and cell sheet formation were observed under an inverted microscope.RESULTS AND CONCLUSION:Dog BMSCs following 24 hours of primary culture presented ellipse or polygonal shape.Most cells adhered at hour 72,and cell colonies were visible at day 7.Cells showed long spindle and completely confluence at day 12,with unclear boundary.BMSCs in the temperature-responsive culture dishes presented short spindle shape,and gradually separated from the dish bottom,forming entire cell sheet containing extracellular matrix at 20 V.These verified that dog BMSCs can be effectively obtained with method of density gradient centrifugation.Complete cell sheet layer can be fabricated with temperature-responsive culture dishes.

We investigated the development of an injectable, biodegradable hydrogel composite of poly(trimethylene carbonate)-F127-poly(trimethylene carbonate)(PTMC11-F127-PTMC11 )loaded with bone morphogenetic protein-2 (BMP-2) derived peptide P24 for ectopic bone formation in vivo and evaluated its release kinetics in vitro. Then we evaluated P24 peptide release kinetics from different concentration of PTMC11-F127-PTMC11 hydrogel in vitro using bicinchoninic acid (BCA)assay. P24/ PTMC11-F127-PTMC11 hydrogel was implanted into each rat's erector muscle of spine and ectopic bone formation of the implanted gel in vivo was detected by hematoxylin and eosin stain (HE). PTMC11-F127-PTMC11 hydrogel with concentration more than 20 percent showed sustained slow release for one month after the initial burst release. Bone trabeculae surround the P24/ PTMC11-F127-PTMC11 hydrogel was shown at the end of six weeks by hematoxylin and eosin stain. These results indicated that encapsulated bone morphogenetic protein (BMP-2) derived peptide P24 remained viable in vivo, thus suggesting the potential of PTMC11-F127-PT- MC11 composite hydrogels as part of a novel strategy for localized delivery of bioactive molecules.
Animals ; Biocompatible Materials ; chemistry ; Bone Morphogenetic Proteins ; pharmacology ; Bone and Bones ; drug effects ; Dioxanes ; chemistry ; Drug Delivery Systems ; Hydrogels ; chemistry ; Osteogenesis ; drug effects ; Peptides ; Prostheses and Implants ; Rats