Objective To identify energy metabolism-proteins of mice midbrain by using the proteomic technique,and to investi-gate the relationship between these proteins and neural diseases.Methods Two-dimensional gel electrophoresis was used to sepa-rate totally soluble proteins extracted from mice midbrain.Some protein spots on two-dimensional gel electrophoresis gels were ana-lyzed by using matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).Results 24 protein spots related with energy metabolism were separated by two-dimensional gel electrophoresis and identified by using MALDI-TOF MS successfully.Conclusion The establishment of energy metabolism-related proteins map of mice midbrain lays a foundation for the research on the involvement of these proteins in neural disease pathogenesis.

Objective In order to assess the value of Echinoccocus granulosus cyclophilin A (EgypA) in immune diagnosis,this novel gene was cloned and expressed.Methods By screening the EST library,the coding region of EgCypA was identified,and the PCR primers were designed based on this sequence.Bioinformatic tools were used to deduce the amino acid sequence of EgCypA and analyzed its biological characteristics.EgCypA was amplified from E.granulosus cDNA library by using PCR.Then,it was cloned into the prokaryotic expression vector pET28a and transformed into E.coli BL21.The recombinant protein was expressed in E.coli BL21 after IPTG induction.The immunogenicity of EgCypA was evaluated by Western blot using the Echinoccocus granulosus and other parasites infected animals' sera.Results Results of SDS-PAGE electrophoresis show that the recombinant BL21 expressed 18 400-25 000 protein,which was identical with the molecular weight calculated by bioinformatic analysis.Western blot shows that the recombinant protein only reacted with its immune serum and E.granulosus cystic fluid immune serum,and EgCypA immune serum could react with the excretion and secretion antigen from E.granulosus protoscolexes,and no cross-reaction between EgCypA and sera from other parasites infected animals.Conclusions Cloning and expression of EgCypA are successful.EgCypA has good immunologic activity and could be a candidate molecular for immune diagnosis of echinococcosis in early stage.

Objective To assess the patient effective dose during SPECT/CT imaging.Methods A total of 157 patients (85 males,72 females;mean age:(60.46±13.44) years,range:24-88 years) who underwent SPECT/CT from April 2013 to September 2014 were retrospectively analyzed.They were divided into 3 groups:99Tcm-MDP bone scan group (n=113),99Tcm-MIBI parathyroid imaging group (n=23) and 99Tcm-MAA pulmonary perfusion imaging group (n=21).The injection dose (MBq) was recorded and the effective dose (mSv) was estimated according to the International Commission on Radiological Protection (ICRP) guideline.The volume-weighted computed tomography dose index (CTDIvol) and dose length product (DLP) of CT were also recorded and the effective dose (mSv) due to CT was calculated.The effective doses among different groups were statistically compared by Kruskal-Wallis rank sum test and Wilcoxon rank sum test.Results The effective dose from radiopharmaceuticals were (4.02±0.35) mSv for 99Tcm-MDP bone scan,(6.13±0.53) mSv for 99Tcm-MIBI parathyroid imaging and (1.66±0.13) mSv for 99Tcm-MAA pulmonary perfusion imaging (x2=96.853;Z =7.533,7.262 and 5.676;all P＜ 0.05).The effective dose due to CT were (3.84±1.98),(1.04±0.19),(4.05±0.47) mSv respectively (x2 =41.225,P＜0.05),and there were significant differences between 99Tcm-MIBI parathyroid imaging group and 99Tcm-MDP bone scan group,99Tcm-MAA pulmonary perfusion imaging group(Z=6.046 and 5.672,both P＜0.05).The total effective dose during SPECT/CT imaging were (7.86± 1.98),(7.21±0.81) and (5.71±0.45) mSv (x2 =28.988,P＜ 0.05).There were significant differences between 99Tcm-MAA pulmonary perfusion imaging group and 99Tcm-MDP bone scan group,99Tcm-MIBI parathyroid imaging group(Z=4.967 and 4.994,both P＜0.05).Conclusions The increase in effective dose due to spiral CT during SPECT/CT examination should be concerned.Appropriate utilization and optimization of SPECT/CT is needed.

Objective To study the effect ofTiaomai-Yin with the therapy method of clearing heat and cooling blood to treat the tachyarrhythmia with the TCM patterns of deficiency of Qi and Yin, and heat of blood stasis.MethodsA total of 60 patients with tachyarrhythmia were randomly recruited into a control group and a treatment group, 30 patients in each. The treatment group was treated withTiaomai-Yin, while the control group was treated with Metoprolol. After both groups were treated for one month, the changes of TCM symptoms, ECG and 24 h DCG were observed.Results The total effective rate of the treatment groups was significantly better than the control group(86.67%vs. 56.67%;χ2=7.957,P<0.05).Tiaomai-Yin significantly improve the symptoms of shortness of breath (1.40 ± 2.11vs. 2.10 ± 1.87;t=-3.341,P<0.01), tiredness (1.30 ± 2.33vs. 2.10 ± 1.84;t=-3.262,P<0.01), dry mouth (0.09 ± 1.34vs. 2.50 ± 2.41;t=-3.194,P<0.01), and tongue (1.90 ± 1.65vs. 3.90 ± 1.81;t=-3.217,P<0.01) and pulse (2.60 ± 1.71vs. 5.10 ± 2.55;t=-3.283,P<0.01).ConclusionTiaomai-Yin had effects in treating tachyarrhythmia with the TCM patterns of deficiency of Qi and Yin, and heat of blood stasis with improvement of symptoms and fewer side effects.

Objective To investigate the anti-tumor activity of triterpenoids from Ganoderma lucidum on different tumor cells . Methods The MTT assay was adopted to detect the in vitro inhibition effect on 5 kinds of tumor cells .The inhibiting curve was drawn ,IC50 was calculated for reflecting the compound′s cytotoxic activity .Results The in vitro experiments demonstrated that three kinds of triterpenoids compound monomer showed different degrees of inhibition effect ,in which the inhibitory effect of gano-derenic acid Y was stronger ,its IC50 on H460 lung cancer cells was 22 .4 μmol/L ,followed by 7-oxo-ganoderic acid Z2 ,its IC50 was 43 .1 μmol/L .Conclusion Ganoderenic acid Y shows a strong inhibitory activity on H 460 lung cancer cells ,7-oxo-ganoderic acid Z2 shows a certain inhibitory activity on H 460 lung cancer cells ,moreover the inhibitory activity is dose dependent .The three com-pounds of ganoderenic acid Y ,7-oxo-ganoderic acid Z2 and ganoderon B have no activity or very weak activity to the other detected cell lines .The anti-lung cancer activity of ganoderenic acid Y and 7-oxo-ganoderic acid Z2 needs to be further deeply studied .

Objective:To establish the lung blast injury model in mice, detect the proteomic changes of lung in mice at different time points, and explore the mechanism of lung blast injury.Methods:A total of 60 healthy male C57BL/6 mice were randomly (random number) divided into the control group, 12-h group after thorax blast, 24-h group, 48-h group, 72-h group and 1-week group ( n=10 each group). Experiments were carried out in the animal laboratory of the General Hospital of the Northern Theater Command. The model of lung blast injury in mice was established by using a self-developed precision blast device, and the lung tissue injury situation was evaluated by gross observation and HE staining. The proteins in mouse lung tissue were quantitatively analyzed based on LC-MS/MS proteomic technology, and the differentially expressed proteins were screened. On this basis, bioinformatics tool was used to analyze proteomic changes. Results:After lung blast injury, scattered bleeding spots could be observed on the surface of lung tissue of mice, and the bleeding points were gradually increased with time, showing a patchy distribution, and the symptoms were the most severe at 24 h. The results of HE staining showed that the normal tissue structure of alveoli disappeared at 12 and 24 h under light microscopy with diffuse bleeding in the alveolar cavity, infiltration of a large number of inflammatory cells, increased interstitial exudate, thickened alveolar wall, and collapsed and merged alveolar cavity. A total of 6 861 proteins were identified by LC-MS/MS in lung tissue samples of mice after thorax blast, and 608 differentially expressed proteins were quantified, of which 227, 140, 202, 258 and 71 differential proteins were at 12 h, 24 h, 48 h, 72 h, and 1 week, respectively. According to GO analysis, 130 biological process subtypes including cell adhesion, extracellular matrix tissue and collagen fibril tissue were obtained. Besides, 66 cellular component involving extracellular exosomes, extracellular matrix and cytoplasm were obtained. And 43 molecular functional subclasses such as extracellular matrix structure composition, actin binding and antioxidant activity were obtained. KEGG analysis yielded 24 pathways including ECM-receptor interactions, focal adhesions and PI3K-Akt signaling pathway across the endothelium.Conclusions:Differentially expressed protein combinations are also different at different time points in the early stage after lung blast in mice, and the injury mechanism is complicated. The lung blast injury is the most serious at 12-24 h after blast and produces significant inflammatory response.

BACKGROUND:Numerous studies have demonstrated that estrogen can regulate the proliferation and migration of endothelial progenitor cel s (EPCs), while EPCs can also promote the function and activity of bone marrow mesenchymal stem cel s (BMSCs) in vitro. OBJECTIVE:To evaluate the ability of the BMSCs and EPCs which construct the composite cel sheet in the repair of alveolar bone defect in ovariectomized rats. METHODS:BMSCs/EPCs composite sheet, EPCs sheet and BMSCs sheet were respectively implanted into the defects of the alveolar bone in ovariectomized rats. Rats with no implantation served as control group. Repaired alveolar bone was assessed by gross examination, histological observation and micro-CT scan at 2, 4, 8 weeks after operation. RESULTS AND CONCLUSION:BMSCs/EPCs composite sheet has greater osteogensis activity and bone repair capacity than BMSCs or EPCs sheet alone.

AIM:To explore the influence of exogenous hydrogen sulfide ( H2 S) on coagulation and fibrinoly-sis in ferric chloride ( FeCl3 )-induced mouse carotid artery thrombosis .METHODS: The mice were divided into sham control group, model group, different concentrations (12.5, 25 and 50μmol/kg) of sodium hydrosulfide (NaHS, H2S do-nor) groups and 30 mg/kg clopidogrel ( positive control ) group.Intraperitoneal injection of NaHS at different concentra-tions and oral administration of clopidogrel bisulfate were performed for 3 d prior to FeCl 3-induced carotid artery thrombo-sis.The frozen sections of the carotid artery were collected to perform HE staining , and the thrombus pattern and the chan-ges of vascular pathology were observed .The thrombus was weighed to calculate thrombosis inhibitory rate .Prothrombin time ( PT) , activated partial thromboplastin time ( APTT) , fibrinogen ( FIB) and fibrinogen degradation product ( FDP) in the mice were also measured by a coagulometer .The plasma levels of thromboxane B 2 ( TXB2 ) , 6-keto-prostaglandin F 1α(6-keto-PGF1α) and plasminogen activator inhibitor (PAI) were detected by ELISA.RESULTS: Compared with model group, NaHS dose-dependently inhibited the formation of carotid artery thrombus .NaHS treatment reduced the contents of TXB2 and PAI, and recovered 6-keto-PGF1αcontent in thrombosis model group .In NaHS treatment groups , 6-keto-PGF1α/TXB2 and thrombus weight was negatively correlated .NaHS treatment prolonged PT and APTT , reduced the content of FIB, but increased the level of FDP in thrombosis model group .CONCLUSION:Hydrogen sulfide prevents FeCl 3-induced carotid artery thrombosis by inhibiting coagulation and activating fibrinolysis .

Objective Patients with aortic dissection have a significant incidence of coronary artery disease.The purpose of this study is to evaluate the safety and feasibility of percutaneous coronary stent in patients who have undergone endovascular stent,and to assess the effect of anti-coagulant and anti-platelet treatment on patients' thrombosis process.Methods From January 2005 to July 2007,8 patients who had undergone endovascular stent-graft during the past 1 to 7 months for type B aortic dissection repair,underwent percutaneous coronary intervention (PCI) because of coexisting coronary artery disease.Anti-coagulant and anti-platelet treatments were administrated after PCI according to the standard protocol.Patients were followed up for a mean period of 23 months.Clinical and false lumen status data were collected during the follow-up.Results PCI were technically successful in all 8 patients and no severe complications such as death,paraplegia,renal failure occurred during hospitalization.Complete false lumen thrombosis was observed in 5 patients and incomplete false lumen thrombosis in the remained 3 patients at the end of follow up.There were no major complications such as death,dissection rupture or aneurysm development occurred during the follow-up period.Conclusion Our data implied that PCI can be safely performed in patients with type B aortic dissection who have undergone endovascular stent-graft,without interrupting the thrombosis process.

Objective To study the genetic diversity in Fujian Sanming Sarcandra Glabra from different geographical provenances;To construct genetic relationship diagram.Methods DNA in leaves was extracted by CTAB. Analysis and evaluation of DNA molecular level of 45 samples of different geographical provenances were conducted by ISSR method. POPgen32 software was used to calculate the genetic diversity and establish gene trees. NTSYS software was used to carry out cluster analysis.Results Six ISSR primers amplified 630 bands. Genetic diversity analysis showed that the average effective number of alleles of 45 varieties was 1.620 2;the average Nei's genetic diversity index was 0.364 5;the average Shannon information index was 0.543 2. Each point had different levels of genetic diversity. Nei's genetic diversity index of the maximum value was 0.497 2, and the minimum value was 0.107 8;Maximum Shannon information index was 0.690 3, and the minimum value was 0.219 2. Cluster analysis results showed that 45 varieties and 14 loci band data were the primitive matrix. 630 genetic similarity coefficients between two different species were obtained. The maximum similarity coefficient among different groups was 1.0, and the minimum was 0.516.Conclusion Different varieties of Fujian Sanming Sarcandra Glabra exist abundant genetic variation and has the molecular basis of abundant species. Using 0.610 as the threshold value can divide Sarcandra Glabra from 45 different geographical provenances into 6 groups. The genetic distance and geographical distance was not related.