Objective To investigate the association between the expression of heparanase(Hpa) and the invasion of choriocarcinoma by studying the expression of Hpa in human choriocarcinoma cell lines JEG-3 and JAR and human chorionic villous tissues.Methods(1)Matrigcl invasion assays were used to detect in vitro invasive ability of JEG-3 cells and JAR cells.(2)Expression of Hpa protein in the human chorionic villous tissues and choriocarcinoma cell lines(JEG-3 cells and JAR cells)were detected by immunocytochemistry and western blot.Results(1)The invasive cell number was significantly larger in JEG-3 cells than in JAR cells(191?17 vs 106?13,P

Intracytoplasmic sperm injection,which combined with in vitro fertilization and micromanipulation techniques,has been applied to research the molecular mechanisms of fertilization,and produce the sexing livestock and transgenic animals.The research advances in porcine intracytoplasmic sperm injection,including in vitro maturation and pretreatment of oocytes,selection and treatment of spermatozoon,artificial activation of injected oocytes after injection,and improvement of the operation technique were reviewed.

Objective To investigate the effect,security and tolerance of pregabalin combined with sertraline hydrochloride for refractory neuropathic pain.Methods Forty-six patients with refractory neuropathic pain were divided into two groups by random digits table.The patients in pregabalin group (24 patients) were treated by pregabalin combined with sertraline hydrochloride,and the patients in carbamazepine group (22 patients) were treated by carbamazepine combined with sertraline hydrochloride.They were treated for 4 weeks.The scores of visual analogue scales (VAS),Hamilton rating scale of depression (HAMD),Hamilton rating scale for anxiety (HAMA) and the dose,adverse reaction were compared between two groups.Results The total effective rate of pregabalin group was 83.3％(20/24) and carbamazepine group was 86.4％ (19/22),there was no significant difference (P＞ 0.05).After treatment of 1,2,4 weeks,the scores of VAS in two groups were significantly decreased compared with those before treatment[pregabalin group:(5.90 ± 1.11),(4.59 ± 1.33),(3.59 ± 1.94) scores vs.(6.50 ± 0.85) scores; carbamazepine group:(5.50 ± 1.06),(4.18 ± 1.71),(3.73 ± 1.88) scores vs.(6.50 ± 0.80) scores] (P ＜0.05).After treatment of 2 weeks,the scores of VAS in carbamazepine group was significantly lower than that in pregabalin group (P ＜ 0.05).In pregabalin group,the scores of HAMD/HAMA after treatment of 1 week had no significant difference compared with before treatment (P ＞ 0.05),and the scores of HAMD/HAMA after treatment of 2,4 weeks were significantly increased [(15.25 ± 4.15),(11.29 ± 4.16)scores vs.(16.08 ± 3.56) scores and (14.21 ± 4.72),(11.63 ± 4.50) scores vs.(15.25 ± 4.05) scores] (P ＜0.05).In carbamazepine group,the scores of HAMD/HAMA after treatment of 1,2 weeks was decreased compared with those before treatment,but there was no significant difference (P ＞ 0.05).The scores of HAMD/HAMA after treatment of 4 weeks was significantly decreased [(13.18 ± 2.42) scores vs.(15.20 ±1.05) scores,(12.32 ± 2.64) scores vs.(14.55 ± 4.43) scores] (P ＜ 0.05).The scores of HAMD/HAMA in pregabalin group after treatment of 4 weeks was significantly lower than that in carbamazepine group (P ＜0.05).There was no severe adverse reaction.Conclusion Pregabalin combined with sertraline hydrochloride is worth to recommend for refractory neuropathic pain,and has no serious adverse reaction.

AIM To investigate that Danhong Injection may decrease inflammation and oxidative stress response in rat peritoneal mesothelial cells (RPMCs) cultured with high glucose.METHODS The third passage of RPMCs were used for the experiment.After being incubated with DMEM for 24 hours,RPMCs were divided into normal control group,high glucose group (RPMCs were incubate with 100 mmol/L glucose for 12 h,24 h,48 h,72 h),Danhong Injection group (complete medium with 80 mL/L Danhong Injection for 48 h) and high glucose + Danhong Injection group (100 mmol/L high glucose with 40 mL/L,80 mL/L,160 mL/L Danhong Injection for 48 h respectively).The cell proliferation was detected by MTT method.The levels of interleukin-18 (IL-18),interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in culture fluid were determined by enzyme linked immunosorbent assay.The contents of malondialdehyde (MDA),superoxide dismutase (SOD),glutathione (GSH),in the culture fluid and intracellular reactive oxygen species (ROS) were measured by kits.The expressions of Bcl-2,Bax,fibronectin (FN),endothelin-1 (EDN1) and haem oxygenase-1 (HMOX1) mRNA were detected by quantitative real-time PCR.The protein expressions of EDN1 and HMOX1 were measured by Western blot.RESULTS High glucose can significantly inhibit RPMCs' viability.High glucose can also up-regulate the expressions of IL-18,IL-6,TNF-α,ROS,MDA,Bax mRNA,FN mRNA,EDN1 mRNA and down-regulate the levels of SOD,GSH,Bcl-2 mRNA,HMOX1 mRNA in RPMCs.Treatment with Danhong Injection can effectively reverse aforementioned effect induced by high glucose.CONCLUSION Danhong Injection can reverse the inhibition of cell viability,the inflammation and oxidative stress response induced by high glucose in RPMCs,thus protect peritoneal membrane.

Animals ; Astragalus Plant ; Cardiomyopathies ; Myocardial Ischemia ; Myocardium ; Rats

Objective:To establish a method of inducing dendritic cells(DC)from the hematopoitic stem cells of rats in vitro,and to identify the phenotype and fusion of DC. Methods:DC obtained from SD rat bone marrow hematopoitic stem cells were propagated in vitro under the condition of rGM CSF、rIL 4 and nrhTNF ?.DC were purified by monoclonal antibody OX62 and magnetic beads.Then DC harvested 12 d later were identified by morphological features,surface antigen expressions and the ability to atimulate T cells. Results:After culture and induction,DC displayed typical morphology with elongated dendritic processes viewed by inversion microscope as well as electron microscope.DC expressed high level surface antigens,including OX62 62.19%;MHCⅠ 70.40%;MHCⅡ 78.28%;CD80 55.58%; CD86 68.38%, The results of mixed lymphocyte reaction(MLR)showed that DC had the ability to stimulate vigorous proliferation of allo T cells. Conclusion:Matue DC could be generated from rat bone marrow hematopoitic stem cells,which presents the feasibility for further clinical application of DC in the immunotherapy of cancers.

Objective To investigate the specific antitumor effects induced by allogeneic dendritic cells (DCs)/osteosarcoma cell fusion vaccine in rats. Methods Fusion vaccine of DCs derived from Wistar rat bone marrow and osteosarcoma cells (UMR106) derived from SD rat was generated by electroporation method, and purified by immunomagnetic beads coated with monoclonal antibody OX62, then cocultured with T lymphocytes derived from SD bone marrow to stimulate the proliferation of the T lymphocytes. The proportion of CD8+ and 44+ cells was determined by flow cytometry, and the anti-tumor activity of cytotoxic T lymphocytes (CTLs) was determined by MTT assay. The SD rat osteosarcoma model was established and active immunotherapy was performed through intradermal injection, and the survival rate of the model rats was observed. Results After cocultured with allogeneic fusion vaccine, the proliferation of T cells increased significantly, and the proportion of CD8+ cells increased from 34.2% to 74.9%, while of CD4+ cells decreased from 59.2% to 19.1% (P

Objective To analyse the flow characteristics in the true lumen and false lumen,and the relationship between the flow characteristics and the collapse degree of the true lumen using MRI.Methods Eleven patients with acute Stanford type B aortic dissection were examined by true FISP、3D CE MRA、PC cine MRI on a Siemens Sonata 1.5 T.Not only the quantitative data on the hemodynamics such as peak velocity,average velocity,average flow volume,forward volume,retrograde volume and net volume,and the area of the true lumen and false lumen can be acquired,but also the blood flow model,ie the velocity-mapping.Then we analysed the relationship between the flow characteristics and the collapse degree of the true lumen.Results The average area of the false lumen in the proximal descending aorta(about 2 cm distal to the entry)was(8.10±2.93)cm2,and(2.59±0.93)cm2 of the true lumen in the same slice (P＜0.05).The average velocity in the false lumen,(2.81±0.73)cm/s,was significantly lower than in the true lumen[(15.52±2.84)cm/s,P＜0.01],wheras the average flow(36.32±5.37)ml/s,was not significantly difierent(P＞0.05)from the average flow in the false lumen(37.62±24.58)ml/s.The velocity-mapping curve looked like same in the true and false lumen in this level.And in the abdominal aorta(about the level of the hepatic hilar),the average flow(10.46±5.57)cm/s was significantly lower(P＜0.05)than in true lumen[(4.04±2.96)cm/s].At this level,the direction of blood flow in the true lumen was retrograde(upward)in the mid and late systolic phase in six patients,and normal in the diastolic phase and early systolic phase,that was to say,bidirectional blood flow can be caught in the true lumen of the abdominal aorta.The collapse degree of the true lumen was closely correlated with the the average velocity and the flow volume in the false Iumen,and the coefficient correlation and P value were 0.931 and 0.000,0.926 and 0.000 respectively.Conclusions PC cine MRI can quantitatively measure the peak velocity,average velocity,average flow volume,forward volume,retrograde volume and net volume,and combined with 3D CE MRA can evaluate the collapse degree of the true lumen.It is important for clinical application in the diagnosis,therapeutic management and the therapeutic opportunity choice of the acute Stanford type B aortic dissection.

Objective To evaluate accuracy and consistency of the coagulation convention that involves PT, INR, FIB, APTT on two coagulation analysators(ACL-9000 and ACL-200, Beckman-Coulter Corporation, US). Methods The repeatability test was used to assess the precision, the accuracy and consistency of the coagulation convention on two coagulation analysators were evaluated by comparison and analysis. Results The repeatability of related test results based on ACL-9000 coagulation analysator was better that CV value less than 5%, but the repeatability of abnormal specimens FIB based on ACL-200 was poor that CV value of 6.6%. Through comparing ACL-9000 analyzer, the test results of ACL- 200 coagulation analysator has good correlation with it, correlation coefficient ?﹥0.975. Conclusion In the condition of the excellent precision and stability, we can also find system errors existed in the analyzer on time by verificating. Through being adjusted and calibrated, the examination department can deliver the results of accuracy consistency.

Objective To investigate the expression and significance of signal pathway Wnt/β-catenin in prostate cancer stem cells.Methods Prostate cancer tissues,hyperplasia prostate tissues and normal prostate tissues were collected,then prostate cancer stem cells were selected from cell suspension in the culture system of serum-free medium by magnetic activated cell sorting system.Immunohistochemical SP test,RT-PCR and Western blot were applied to test the expression of Wnt and β-Catenin mRNA or protein in prostate cancer stem cells,hyperplasia prostate tissues and normal prostate tissues.Results The protein expression of Wnt and β-Catenin was higher in prostate cancer tissues compared with that in hyperplasia prostate tissues and normal prostate tissues;mRNA expression of Wnt and β-Catenin was higher in prostate cancer stem cells (4.57±0.83,3.93±0.78) than in hyperplasia prostate tissues (1.32±0.35,1.48±0.44) and normal prostate tissues (1.00±0.12,1.00±0.11),and the difference was statistically significant (F＝13.287,12.648,P＝0.000).Protein expression of Wnt and β-Catenin was higher in prostate cancer stem cells(0.87±0.10,1.12±0.23) than in hyperplasia prostate tissues(0.39±0.08,0.64±±0.11) and normal prostate tissues (0.33±0.09,0.45±0.10),and the difference was statistically significant (F＝16.625,14.417,P＝0.000).Conclusion Signal pathway Wnt/β-catenin is stimulated abnormally in prostate cancer stem cells,causing the occurrence of prostate cancer,providing a new research direction for treatment of prostate cancer.