Objective To investigate the optimal time points for transplantation with in vitro cultured dopaminergic neurons as a potential treatment of Parkinson’s disease. Methods After the ventral mesencephalic precursors from E11 rat embryos were expanded for 7 days in vitro, they were harvested respectively from day 0 to day 7 in the period of induced differentiation. Then the cells were seeded again and cultured for another 7 days. The final survival of dopaminergic neurons determined the optimal time points for moving cells in vitro and, possibly, for transplantation in vivo. Result The survival rate of dopaminergic neurons moved on day 0 and day 1 in the period of induced differentiation was significantly higher than that at other time points. Conclusion The optimal time points for transplantation are found possibly to be day 0 and day 1 in the period of differentiation after the expansion of ventral mesencephalic precursors from E11 rat embryos.

Objective To prepare the quantum dots(QDs) (CdTe-MPA)-loaded polymer(lactic-coglycolic acid,PLGA) microbubbles(MBQDs@PLGA) as dual-modality imaging agent for both fluorescent and ultrasonic imaging ability.Methods The MBQDs@PLGA were generated by the double emulsion technique,then filling in C3F8 after freeze-drying.Confocal laser scanning microscope(CLSM) and transmission electron microscope(TEM) were used to confirm the load of quantum dots in the MBs.Fluorospectro photometer spectra of the MBQDs@PLGA were analyzed to demonstrate the fluorescent imaging ability and determine the encapsulation efficiency by using the regression equation.Imaging experiments was applied to validate the fluorescent and ultrasonic imaging ability of the MBQDs@PLGA both by imaging of the model in vitro and by imaging of ovarian tumor blood vessels of tumor-bearing nude mouse in vivo.Results At excitation 272 nm the MBQDs@PLGA peak of the emission spectrum was 549 nm,and the encapsulation efficiency was 54％.The average diameter of MBQDs@PLGA was (1.7 ±0.2)μm,CLSM and TEM results confirmed the QDs-loaded in MBQDs-PLGA.The imaging results of MBQDs@PLGA showed a dual-modality imaging ability both fluorescent and ultrasonic imaging.Conclusions MBQDs@PLGA present fluorescence-ultrasound dual mode imaging performance by the QDs embedding in polymer microbubbles,and explore a new development train of thought of multi-mode imaging agent.

Objective To investigate the effect of Yunnan Baiyao Plaster on serum related markers levels in the patients with stroke ,to evaluate its improvement effect on the neurological dysfunction and pain and to analyze its effect mechanism .Methods Eighty cases of hemorrhagic stroke treated in the neurosurgical department of our hospital from February 2015 to June 2016 were screened as the research subjects and divided into the control group and observation group according to the random number table method ,40 cases in each group .All subjects were given the routine treatment ,while the observation group received the external use of Yunnan Baiyao Plaster around the head operative incision and drilling ,to give Yunnan white ointment ,with the attaching area of 5 cm × 4 cm ,once daily ,for consecutive 7 d ,meanwhile took oral Yunnan Baiyao Plaster 0 .5 g with warm boiling water ,once daily for consecutive 7 d .The serum markers levels before and after treatment ,neural function defect score ,decline proportion of NIHSS scores and prognostic indicators were compared between the two groups .Results The mean pain value within 1 week in the obser-vation group was lower than that in the control group ,the difference was statistically significant (P<0 .05);the pain peak value and valley value within 1 weeks had no statistical difference between the observation group and control group ,the difference was not statistically significant (P> 0 .05);after 2 weeks ,ET ,TNF-α and IL-1β levels in the observation group and control group were lower than those before treatment in the intra-group comparison ,while the comparison between the two groups showed that the ob-servation group was lower than the control group ,the difference was statistically significant (P<0 .05);the decrease rate of NIH-SS scores within 2 weeks in the observation group was (74 .4 ± 5 .2)% ,which was higher than (66 .3 ± 10 .3)% in the control group ,the difference was statistically significant (P<0 .05);the occurrence rate of severe complications within 2 weeks in the ob-servation group was 7 .5% (3/40) ,the mortality rate was 2 .5% (1/40) ,which in the control group were 12 .5% (5/40) and 2 .5%(1/40) ,the difference was not statistically significant (P>0 .05) .Conclusion The combined treatment of Yunnan Baiyao Plaster in the patients with hemorrhagic stroke conld relieve pain and reduce the neurological deficit ,which might be related to the mechanism of anti-inflammation and analgesia .

Objective To quantify the right ventricular (RV) function in patients with left heart disease (LHD) related pulmonary hypertension (PH) by two-dimensional speckle tracking imaging (2D-STI),and explore its potential value in diagnosis of reactive PH.Methods Forty healthy volunteers and 65 patients with LHD related PH were enrolled.PH patients were further divided into two subgroups according to pulmonary vascular resistance(PVR):passive PH group and reactive PH group.A complete 2D-STI derived longitudinal strain of RV free wall was performed,and the basal,middle,apical longitudinal peak systolic strain (LS) and RV free wall global LS (RVLSbas,RVLSmid,RVLSapi and RVLSfw) were measured.Results Compared to control group,RVLSbas and RVLSfw in reactive PH group decreased significantly (all P ＜0.05).Compared to passive PH group,RVLSbas and RVLSfw in patients with reactive PH decreased significantly (all P ＜0.05).The differences of RVLSbas and RVLSfw between control group and passive PH group had no statistical significance (all P ＞0.05).No significant differences in etiology and left ventricular ejection fraction (LVEF) were observed between PH subgroups (all P ＞0.05).The area under the ROC curve of RVLSbas in predicting reactive PH was 0.93,the cut-off value was-21.86,and the sensitivity and specificity were 83 ％ and 89 ％,respectively.The area under the ROC curve of RVLSfw was 0.79,the cut-off value was-13.84,and the sensitivity and specificity were 59％ and 89％,respectively.Conclusions 2D-STI can realize quantitative assessment of RV function in patients with LHD related PH.RVLSbas and RVLSfw may have important clinical value in diagnosing patients with reactive PH,and differential diagnosis value between patients with passive PH and patients with reactive PH.

Purpose To explore the apoptosis mechanism of human ovarian cancer cells SKOV3 processed with ultrasound (US) combined with photodynamic therapy (PDT), with self-contained lactic acid/glycolic acid (PLGA) microbubbles (MBQDs/PLGA) containing quantum dots (QDs) as a photosensitizer. Materials and Methods MBQDs/PLGA was prepared with double emulsion method, and MTT was used to compare the cytotoxic activity difference between QDs and MBQDs/PLGA, to determine the proper treatment condition. SKOV3 cells were treated under selective conditions, cell damage manifestation was observed with HE staining, and double staining flow cytometry was used to detect cell apoptosis. Results When cells were treated with PDT with energy density of 180.0 J/cm2 and US with sound intensity of 0.5 W/cm2 (1.0 MHz, 10 s), cell deformation could be observed under a microscope in both US-PDT-MBQDs/PLGA group and PDT-MBQDs/PLGA group, and cell connections were absent between cells;transmission electron microscope showed dense chromatin gathered along the nuclear membrane, with the formation of apoptotic bodies also be displayed. Maximum apoptosis rate was (22.17±0.38)% Conclusion MBQDs/PLGA-mediated PDT contains proliferation inhibition effect for SKOV3 cells, which can be synergied with low-power ultrasonic irradiation due to sonoporation effect.

Objective To explore an effective evaluation method for students' self-and peer-assess-ment. Methods The students of 6 groups participating in extracurricular teaching activities were selected as research subject. Traditional method (final score = mean score of group/2 + teacher's score/2) and mean difference method [final score=teacher's score-(mean difference of group-mean difference of all groups)] were used to calculate final score of each group, and effect of two methods were compared. Results Scores of most groups were higher than the teacher's scores, and high scores were given by group 3 in self- and peer-assessment. The final score of all groups were higher than teacher's scores in traditional method. Compared with teacher's scores, final scores increased significantly in group 1, 4, 5 below mean difference, final score decreased significantly in group 2, 3 above mean difference, and final score did not differ in group 6 equal to mean difference in mean difference method. Conclusion The mean difference method can reflect the effect of student's self- and peer-assessment, and guide student to make objective and accurate evaluation. It is a more reasonable and scientific evaluation method for self-and peer-assessment.

BACKGROUND:A series of studies have demonstrated that ventral mesencephalic dopamine precursors may differentiate into considerable dopaminergic neurons (DNs) available for cell transplantation for Parkinson's disease.However,the detailed molecular characteristics of these cells remain undear.OBJECTIVE:To observe the expression of main marker genes of DNs cultured in vitro.DESIGN,TIME AND SETTING:This observation experiment was performed at the Brain Tumor Laboratory in the Second Affiliated Hospital of Soochow University and Shanghai Biochip Company from December 2005 to December 2008.MATERIALS:Inbred pregnant Wistar rats (F+36) weighing 300 g approximately were used for in vitro culture of mesencephalic cells.METHODS:Ventral mesencephalic cells from inbred pregnant Wistar rats were cultured with basic fibroblast growth factor,and were induced to differentiate in vitro with L-Ascorbic acid-2-phosphate sesquimagnesium salt.Total RNA were extracted before and 4,7 days after the induction.Affymetrix microarrays were utilized to detect the expression changes of main marker genes of DNs.MAIN OUTCOME MEASURES:Morphologic changes of differentiated ventral mesencephalic precursors;Expression of DN-ralated common marker genes before and after the induced differentiation.RESULTS:Rat ventral mesencephalic precursors differentiated into numerous tyrosine hydroxylase-positive cells,which expressed multiple marker genes of DNs,such as AHD2,calcium binding protein 1 (Cab1),DDC,DAT,Drd2,Girk2,VMAT-2,with variously degrees and with different tendencies.Among those genes,only Cab1 was significantly up-regulated and only Girk2 was significantiy down-regulated.CONCLUSION:Rat ventral mesencephalic precursors can be induced to differentiate into plenty of tyrosine hydroxylase-positive DNs,mainly originating in A10 regions.

Objective: To identify Persicae Semen and its sibling species, and to secure the quality and clinical safety of this medicinal material. Methods: The internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA of Persicae Semen and its sibling species were amplified and bidirectionally sequenced by DNA barcoding. Sequence assembly and consensus sequence generation were performed by the CodonCode Aligner 4.1. The genetic distances were computed by MEGA 5.0 in accordance with the Kimura 2-parameter (K2P) model, and the phylogenetic tree constructed by the neighbor-joining (NJ) method. Results: The length of ITS2 sequences of the two origin plants of Persicae Semen was between 212 bp to 213 bp. Their intraspecific genetic distance was much lower than the interspecific genetic distance with their sibling species. The ITS2 sequence possessed the character of good stability and low intra-specific sequence variation. In the NJ tree, both Prunus persica and P. davidiana were clustered into one large branch, and clearly separated with their sibling species. Conclusion: ITS2 can be used to effectively distinguish Persicae Semen from its sibling species, which can provide a reference for the iden-tification of other Chinese medicine and its sibling species.

Objective: This study aimed to discriminate between Eupatorii Herba and its adulterants in order to guarantee the quality and clinical curative effect of this medicinal material. Methods: Genomic DNA extracted from Eupatorii Herba was used as templates. The internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA was amplified. Sequence assembly and consensus sequence generation were performed by CodonCode Aligner. The intraspecific and interspecific genetic distances of Eupatorii Herba and its adulterants were computed by MEGA5 and the phylogenetic tree was constructed using the neighbor-joining (NJ) method. Results: The length of ITS2 sequence of Eupatorii Herba was 218 bp. The maximum intraspecific genetic distance (K2P distance) of Eupatorii Herba was 0.0092. The minimum interspecific genetic distance of Eupatorii Herba and its adulterants was 0.024. The NJ trees showed that the ITS2 sequence would be used to identify Eupatorii Herba and its adulterants. Con-clusion: ITS2 sequence was able to identify Eupatorii Herba and its adulterants correctly and it provided a new technique to ensure clinical safety in utilization of traditional Chinese medicines.

Objective To study the targeting ability of aptamer-modified PLGA nanoparticle encapsulated with gold nanorod and liquid perflurocarbon,and observe its mediated ultrasound/photoacoustic dual-modality imaging in vitro.Methods PLGA nanoparticle was prepared using double emulsion technique.Aptamer-modified PLGA nanoparticle was obtained by connecting,then PLGA nanoparticle was conjugated with MUC1 aptamer carbodiimide technique aimed to obtain aptamer-modified PLGA nanoparticle.Targeting ability of aptamer-modified PLGA nanoparticle to MCF-7 tumor cells in vitro was tested by fluorescence microscope.Besides,there were three control groups including PLGA nanoparticle group,aptamer group and HELA cell group.A commercial photoacoustic instrument was used to detect ultrasound/photoacoustic signal.Results A significant number of aptamer-modified PLGA nanoparticles gathered around the MCF-7 cells,however,the specific binding can not be observed in control groups.Both ultrasound and photoacoustic signal intensity of the aptamer-modified PLGA nanoparticles increased significantly when excited with laser.Conclusions The aptamer-modified PLGA nanoparticle was successfully prepared,and had superior targeting ability to MCF-7 cells,Morever,ultrasound/photoacoustic signal emitted the by nanoparticle increased significantly.These properties make the targeted nanoparticle a great potential to become a distinguished uhrasound/photoacoustic dual modality contrast agent,which lays good foundation for further targeted imaging application in vivo.