Objective To construct eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG,and to detect the expression of the plasmid in vitro.Methods hOPG cDNA fragment was extracted from plasmid MGC:29565,amplified by PCR,and inserted into pcDNA3.1(-)vector,then identified by restriction endonuclease digestion and nucleotide sequencing.C2C12 cells were transfected with the plasmid using Lipofectamine 2000.The expression of OPG protein was detected by immunohistochemistry techniques and bone wafer pit assay.Results The constructed recombinant plasmid contained the sequence of hOPG gene.After transfection with the plasmid,active OPG protein could be expressed in C2C12 cells.Conclusion The constructed eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG can express OPG in vitro,which is the basis for further study on the treatment of bone absorption caused by abnormal activity of osteoclasts.

Clinical features and thyroid-stimulating antibody(TSAb)in 32 newly diagnosed patients with Graves' ophthalmopathy(GO)were compared with those in 27 Graves' disease(GD)patients without GO(as control group).All of the patients with GO received intravenous glucocorticoids.The level of serum TSAb in patients with GO was significantly higher than that in patients without GO.TSAb was also associated with the prognosis.It suggests that TSAb seems to be the most active component among the TSH receptor antibodies related to ophthalmopathy and may act as a predictive parameter.

Objective To evaluate the feasibility of inducing choroidal neovascularization(CNV) in Brown Norway(BN) rats by diode laser with wavelength of 810 nm. Methods Thirty-five BN rats were divided into(7 groups.)In each groups,one rat served as control,while the other four recieved retinal photocoagulation by diode(laser) in two eyes,at a spot size of 75 ?m with a duration of 100 ms and power of 140 mW.On 1,3,7,14,21,28 and 56 days after photocoagulation,the formation and nature process of CNV were observed by fundus fluorescein(angiography)(FFA),indocyaninegreen angiography(ICGA) and light microscopy(LM). Results FFA,ICGA and LM demonstrated that CNV began to form on the seventh day after photocoagulation,increased on the fourteenth day,reached the peak on the twenty-first day and kept stable until the fifty-sixth day. Conclusion Diode Laser can successfully induced CNV in BN rats,and it is an ideal animal model for further study.

OBJECTIVETo explore methods and clinical outcomes of microsurgical technique in treating patients with severed palm.

METHODSFrom January 2009 to December 2012,45 patients with severed palm were treated by replantation through microsurgical technique, included 37 males and 8 females, aged from 13 to 45 years old with an average of 25. Postoperative survival rate and evaluation standard of upper limb replantation function posposed by Chinese Medical Association were applied for evaluate clinical outcomes after operation.

RESULTSForty-five patients with severed plam were treated by replantation. Thirty-nine patients (121 fingers) were survived,and survival rate was 87%. All patients were followed up 3 to 15.5 months with an average of 11.5 months. According to evaluation standard of upper limb replantation function posposed by Chinese Medical Association, the total score was 80.27 +/- 1.93, and 27 cases got excellent results, 8 good and 4 poor.

CONCLUSIONThe success of replantation of severed palm depends on grasping operation indication strictly, knowing complexity and local anatomic relationship, debridement completely during operation, making full use of remaining organization, arteriovenostomy through microsurgical technique as early as possible, constructing circulation and repairing injuried nerve rationally.

Adolescent ; Adult ; Female ; Hand Injuries ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Replantation ; Young Adult

AlM: To measure ocular biometric values with the degree of axial myopia and determine the relationship between the differences using the Lenstar 900.
METHODS:Totally 413 myopes (826 eyes) were enrolled in this study and were divided into 3 groups: low myopia (myopia <3. 00 diopters, 104 eyes), moderate myopia ( myopia =3. 00-6. 00 diopters, 500 eyes) and high myopia ( myopia > 6. 00 diopters, 222 eyes ). Central corneal thickness ( CCT ) , aqueous depth ( AD ) , lens thickness (LT), axial length (AL) were measured by Lenstar 900. The parameters were tested using analysis of variance and the relations among SE, AL, LT were analyzed.
RESULTS: There were significant difference both in AL (F=206.16, P<0.01) and AD (F=4.764, P<0.05) and no significant difference both in CCT and LT between these myopia groups. With analysis of Person, the Spherical equivalent ( SE) shows a significantly positive correlation with AL (r=0. 662, P<0. 01) and AD (r=0. 095, P<0. 05), no correlation with CCT and LT. AL shows a significantly positive correlation with AD (r=0. 347, P<0. 01) and CCT (r=0.126, P<0.01), negative correlation with LT (r=-0.265, P<0. 01). LT shows a significantly negative correlation with AD (r=-0. 496, P<0. 01).
CONCLUSlON: Along with the diopters increasing in myopia, the axial length and aqueous depth continue to increase. Spherical equivalent ( SE) shows a significantly positive correlation with AD. AL shows positive correlation with SE, AD and CCT and negative correlation with LT. LT shows a significantly negative correlation with AD.

ObjectiveTo investigate and compare the behavioral changes, neuron loss of hippocampus and mossy fiber sprouting between pilocarpine-induced status epilepticus (SE) model and pentylenetetrazole (PTZ) kindling model in rats.MethodsAfter two different epilepsy models were made, Vedio was adopted to observe the behavioral changes. Nissl staining and Neo-timms' staining were separately used to observe and compare the neuron loss of hippocampus and mossy fiber sprouting in the dentate gyrus (DG) at different time points during epileptogenisis.ResultsNo recurrent spontaneous seizure, no neuron loss and no mossy fiber sprouting were found in PTZ kindling model; whereas obvious neuron loss was found in CA1, CA3 of hippocampus and hilus of DG, and mossy fiber sprouting were found in pilocarpine model in parallel with recurrent spontaneous seizures. ConclusionPTZ kindling model resembles absence epilepsy in human, while pilocarpine-induced status epilepticus model resembles chronic temporal epilepsy in human. Neuron loss and mossy fiber sprouting may play an important role in epileptogenisis. Pilocarpine-induced epilepsy model can be regarded as an ideal chronic temporal epilepsy model.

A method of comprehensive chemical pattern recognition of Atractylodis Rhizoma was established by GC-MS fingerprint, principal component analysis, cluster analysis and discriminant analysis. A DB-wax column (0.25 mm x 60 m, 0.25 microm) with El ion source and 70 V electron multiplier were used for GC-MS analysis. Using principal component analysis, cluster analysis, and discriminant analysis, 15 common peaks of sample fingerprints for chemical pattern recognition research were analysed. The same results were obtained from the fingerprint, principal component analysis and cluster analysis, which could use to distinguish genuine Atractylodes lancea, ungenuine A. lancea and A. chinensis. Thus, this method could be used for the quality control and comprehensive evaluation of Atractylodis Rhizoma.
Atractylodes ; chemistry ; China ; Discriminant Analysis ; Drugs, Chinese Herbal ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods ; Quality Control ; Rhizome ; chemistry

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua ; genetics ; metabolism ; Artemisinins ; metabolism ; Biosynthetic Pathways ; Drugs, Chinese Herbal ; Mixed Function Oxygenases ; genetics ; Oxidoreductases ; genetics ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism

OBJECTIVETo study the regulation of luteolin on spleen cells and sarcoma S180 cells in normal ICR mice.

METHODSSpleen cells and S180 cells were incubated with different concentrations of luteolin (50, 100, 200, and 400 μmol/L). The effect of luteolin on spleen cells and sarcoma S180 cells was determined by MTT assay. The apoptosis was detected using propidium iodide staining flow cytometry. Intracellular reactive oxygen species (ROS) was determined by flow cytometric analysis. Activities of free radicals scavenging were determined by hydroxyl radical and DPPH tests.

RESULTSCompared with the solvent control group, 200 and 400 μmol/L luteolin increased the spleen cells viability (P < 0.05). Luteolin at 100, 200, and 400 μmol/L decreased activities of S180 cells (P < 0.01). The proportion of sub-G1 phase spleen cells was reduced after treated with 200 and 400 μmol/L luteolin (P < 0.05). The proportion of sub-G1 phase S180 cells was elevated after treated with 200 and 400 μmol/L luteolin (P < 0.05). Compared with the solvent control group, levels of intracellular ROS in spleen cells of ICR mice all increased; levels of intracellular ROS in S180 cells all decreased after treated with 50, 100, 200, and 400 μmol/L luteolin (P < 0.05). Luteolin scavenged hydroxyl radical and DPPH in a dose dependent manner.

CONCLUSIONLuteolin had bilateral regulation on viability and apoptosis of spleen cells and S180 cells (promoting the viability of spleen cells, inhibiting apoptosis of spleen cells, inhibiting the viability of S180 cells, and promoting apoptosis of S180 cells), which was worth further study and exploration.

Animals ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cell Survival ; Luteolin ; metabolism ; Mice ; Mice, Inbred ICR ; Reactive Oxygen Species ; Sarcoma ; Spleen ; metabolism

The paper discusses the acquisition and utilization of APP medical resources from medical literature,medical information,evidence-based medicine,medical community,and other specialized and various supporting tools,so as to provide references for continuous education and learning of medical staffs.