Abstract:Vascular smooth muscle cells have attracted considerable interest as a model for a flexible program of gene expression. This cell type arises throughout the embryo body plan via poorly understood signaling cascades that direct the expression of transcription factors and microRNAs which, in turn, orchestrate the activation of contractile genes collectively defining this cell lineage. The discovery of myocardin and its close association with serum response factor has represented a major break-through for the molecular understanding of vascular smooth muscle cell differentiation. Retinoids have been shown to improve the outcome of vessel wall remodeling following injury and have provided further insights into the molecular circuitry that defines the vascular smooth muscle cell phenotype. This review summarizes the progress to date in each of these areas of vascular smooth muscle cell biology.

Abstract:As-moral-assurance-of-application-of-law,judicial-ethics-is-indispensable-to-construction-of-law.-In-view-of-dimensions,performances-of-judicial-ethics-reflect-on-two-levels:-pursuing-judicial-justice-far-from-judicial-corruption-and-faithfulness-to-the-law,and-complying-with-the-law.-In-terms-of-conditions-of-realization,judicial-ethics-requires-not-only-good-personnel-ethics-of-justice-workers-who-count-the-judicial-wisdom-as-the-higher-value,but-also-the-lead-of-the-consensus-of-academia-and-judiciary-value-system.

Abstract:Calcium channel blockers (CCBs) are widely used to treat cardiovascular diseases such as hypertension, angina pectoris, hypertrophic cardiomyopathy, and supraventricular tachycardia. CCBs selectively inhibit the inward flow of calcium ions through voltage-gated calcium channels, particularly Cav1.2, that are expressed in the cardiovascular system. Changes to the molecular structure of Cav1.2 channels could affect sensitivity of the channels to blockade by CCBs. Recently, extensive alternative splicing was found in Cav1.2 channels that generated wide phenotypic variations. Cardiac and smooth muscles express slightly different, but functionally important Cav1.2 splice variants. Alternative splicing could also modulate the gating properties of the channels and giving rise to different responses to inhibition by CCBs. Importantly, alternative splicing of Cav1.2 channels may play an important role to influence the outcome of many cardiovascular disorders. Therefore, the understanding of how alternative splicing impacts Cav1.2 channels pharmacology in various diseases and different organs may provide the possibility for individualized therapy with minimal side effects.

Abstract:The-feasibility-of-the-coordinated-and-balanced-development-of-science-and-technology-in-life-sciences-and-bioethics-were-discussed-in-this-article-through-the-exploration-of-the-origin-of-ethical-issues-caused-by-the-development-of-modern-science-and-technology-in-life-sciences,and-the-methodology-was-proposed-for-the-ethical-management-of-the-development-of-life-sciences,hoping-to-find-the-dynamic-balance-concerning-the-development-and-the-ethical-issues-in-life-sciences.

Abstract:In-modern-society,mass-media-has-played-a-very-important-role-in-human-life-with-its-ubiquitous,all-inclusive-and-omnipotent-characteristic.-Under-the-modern-medical-model,strengthening-medical-ethics-social--education-by-using-the-features-of-mass-media-will-have-a-great-impact-on-resolving-the-contradictions-between-patients-and-hospitals,maintaining-social-stability,and-promoting-social-harmony.-Firstly,we-should-take-good-advantage-of-media-to-build-up-a-favorable-medical-ethics-information-environment-and-give-powerful-implements-support-to-medical-ethics-social--education.-Secondly,we-should-train-medical-workers’-media-literacy,improve-their-ability-of-distinguishing-media-information-and-judging-information-value.-Meanwhile,we-should-insist-on-positive-guidance-through-the-cooperation-between-healthcare-and-mass-media,show-model-behaviors-and-set-good-examples-for-medical-presonnel.

Abstract:Objective: MicroRNA (miRNA) expression is deregulated in many types of human cancers. We sought to investigate the expression patterns of the miRNAs, miR-21, miR-145 and miR-155 in sporadic gastric cancer in a Chinese population. Methods: Total RNA was extracted from archived gastric cancer tissues and adjacent non-cancerous tissues from 20 pairs of paraffin-embedded specimens. Expression levels of miR-21, miR-145 and miR-155 were detected by quantitative reverse transcriptase PCR using a specific stem-loop primer, with U6 as the internal reference gene. Results: The expression of miR-21 and miR-155 in gastric cancer samples was significantly higher than in paired non-cancerous samples (P < 0.05). There was no statistically significant difference in expression levels of miR-145 between cancerous and non-cancerous tissues (P > 0.05). Conclusion: In Chinese sporadic gastric cancer tissues, the expressions of the oncogenic miR-21 and miR-155 were significantly up-regulated, while the expression of the tumor suppressor miR-145 was decreased, although this decrease was not statistically significant. Thus there is specificity in the miRNA expression pattern in gastric cancers in the Chinese population.

Abstract:Based-on-knowledge-creation-theory-and-complex-adaptive-system-theory,the-article-researched-on-the-knowledge-sharing-mechanism-of-hospital-and-suggested-the-synergy-managerial-mode-in-order-to-improve-knowledge-sharing-and-service-of-hospital.

Abstract:Objective:-To-learn-the-economic-performance-of-public-medical-institutions-in-Songjiang-district-of-Shanghai-before-and-after-the-separation-of-revenue-and-expenditure.-Methods:-Financial-statements-of-public-medical-institutions-were-collected,and-the-designed-questionnaires-were-filled-by-district-health-bureau.-Results:-After-the-implementation-of-separation-of-revenue-and-expenditure,public-medical-institutions-in-Songjiang-district-had-not-only-gained-steadily-increase-of-financial-assistance,out-patients-and-emergency-patients,-but-also-realized-significanty-decreasined-percent-of-operating-income-and-expense-per-out-patient-or-emergency-patient.-However,public-spending-remained-at-high-level-and-financial-deficit-of-medical-service-still-existed.-Conclusion:-Separation-of-revenue-and-expenditure,which-is-a-kind-of-government-dominating-health-investment-mechanism,has-ensured-good-development-of-public-medical-institutions,facilitated-its-return-to-commonweal,and-made-it-possible-for-implementation-of-basic-drug-system.-However,public-medical-institutions-should-strengthen-budget-management-and-control-operating-cost.

Abstract:Objective: To investigate the effect of cardiomyocyte proliferation induced by human hepatocyte growth factor (HGF) in pigs with chronic myocardial infarction (CMI). Methods: A steerable, deflectable 7F catheter incorporating a 27-guage needle was advanced percutaneously to the left ventricular myocardium of 18 pigs with CMI. Pigs were randomized (1:1:1) to receive adenoviral vector HGF (total dose, 1×1010 genome copies), which was administered as five injections into the infarcted myocardium (total, 1.0 mL), or saline, or Ad-null (control groups). Injections were guided by Ensite NavX left ventricular electroanatomical mapping. HGF and cyclin proteins were detected by western blot and immunoprecipitation analysis. Histological and immunohistochemical analysis determined proliferating cardiomyocytes. Myocardial perfusion and cardiac function were estimated by Gated-Single Photon Emission Computed Tomography (G-SPECT). Results: Western blot analyses showed that HGF were predominantly expressed in the infarct core and border in the myocardium of the infarcted heart. G-SPECT analysis indicated that the HGF group had better cardiac function and myocardial perfusion four weeks after the injection of Ad-HGF than before the injection of Ad-HGF. After treatment there were more proliferating cardiomyocytes in the HGF group compared to either of the control groups. Furthermore, the HGF group myocardial samples expressed higher levels of p-Akt, cyclin A, cyclin E, cyclin D1, cdk2, cdk4 than those in the control groups. Conclusion: The over-expression of HGF activates pro-survival pathways, induces cardiomyocyte proliferation, and improves the perfusion and function of the porcine CMI heart.

Abstract:With-the-quickening-modern-life-pace-and-the-increasing-employment-pressure,more-and-more-people-pay-attention-to-job-burnout.-Because-of-the-professional,the-job-burnout-among-the-medical-staff-is-becoming-more-prevalent-and-serious.-The-article-explored-the-current-situation-and-characteristics-of-occupational-burnout,analyzed-the-related-psychosocial-factors,and-proposed-effective-countermeasure-for-medical-staff-burnout.

Abstract:This paper first applies the sequential cluster method to set up the classification standard of infectious disease incidence state based on the fact that there are many uncertainty characteristics in the incidence course. Then the paper presents a weighted Markov chain, a method which is used to predict the future incidence state. This method assumes the standardized self-coefficients as weights based on the special characteristics of infectious disease incidence being a dependent stochastic variable. It also analyzes the characteristics of infectious diseases incidence via the Markov chain Monte Carlo method to make the long-term benefit of decision optimal. Our method is successfully validated using existing incidents data of infectious diseases in Jiangsu Province. In summation, this paper proposes ways to improve the accuracy of the weighted Markov chain, specifically in the field of infection epidemiology.

Abstract:Developing-community-health-services-is-an-important-function-of-government-to-provide-public-service,and-the-medical-personnel-resources-play-the-vital-role-for-the-quality-of-community-health-service.-The-article-discussed-on-the-condition-of-the-community-health-service-personnel-resources-in-Xuzhou-city-in-order-to-provide-reference-for-the-development-of-community-health-services.

Abstract:Clinic-of-hospital-is-the-place-of-receiving-patients-to-make-general-medicine-and-emergency-handling-which-patients-contact-with-earliest-and-most.-The-hidden-troubles-in-security-of-public-area-of-outpatient-service-concern-the-image-and-status-of-hospital-and-the-whole-public-health-welfare-undertaking.-So-the-medical-guidance-assistant-plays-the-most-direct-and-irreplaceable-role-of-approaching-the-potential-risk-of-public-area-of-clinic.

Abstract:Objective: To knockdown the C-erbB2 gene in salivary gland adenoid cystic carcinoma SACC-83 cells using RNA interference, and determine the effect of silencing C-erbB2 on cell proliferation. Methods: C-erbB2-siRNA was transfected into SACC-83 cells. RT-PCR and immunohistochemistry were used to detect C-erbB2 expression in SACC-83 cells. Cell proliferation was measured by the MTT assay and gene knockdown was achieved by RNA interference. Apoptosis was analyzed by flow cytometry. Results: Compared with the control, C-erbB2 mRNA expression was decreased in the C-erbB2-siRNA transfection group, and immunohistochemical analysis indicated that C-erbB2 protein expression was decreased. After C-erbB2-siRNA was transfected for 48 h, absorbance at 570 nm (MTT) was 0.185±0.021 compared with 0.354±0.034, 0.299±0.053, and 0.314±0.049 in the blank control, liposome control and negative control siRNA groups, respectively. The differences were statistically significant (P < 0.05) between the C-erbB2-siRNA group and the control groups. Following the C-erbB2 knockdown, the percentage of apoptotic cells was 5.63% compared with 2.04%, 2.85%, and 2.98% in the three control groups, respectively. Proliferation of SACC-83 cells was inhibited, and early apoptotic cells were increased. Conclusion: RNA interference can effectively silence C-erbB2 gene expression and inhibit growth of SACC-83 cells, which indicates the potential of targeting this gene as a novel gene therapy approach for the treatment of salivary gland adenoid cystic carcinoma.

Abstract:In-recent-years,-the-increase-of-financial-risks-of-colleges-and-universities-has-arisen-with-the-further-development-of-higher-education-reform-and-the-college-expansion-plan.-The-deterioration-of-financial-situation-influences-the-sustainable-development-of-higher-education.-The-article-discussed-the-importance-of-internal-control-to-financial-risks,-analyzed-the-inner-causes-of-financial-risk,-and-put-forward-some-suggestions-of-improving-internal-financial-controls-in-colleges-and-universities.

Abstract:Objective: To investigate the metabolites of polybrominated diphenyl ether 99 (BDE-99) and its related cytochrome P450s in an in vitro system. Methods: Rat primary hepatocytes were isolated and treated with BDE-99 for 24-72 h. Metabolites were then extracted from the hepatocytes and media, and detected by GC/MS. Several mRNAs of metabolic enzymes were also extracted from the same cells and the gene expression levels were determined using quantitative real-time PCR. In addition, selected recombinant cytochrome P450s (CYPs) were expressed in a bacurovirus/sf9 system, and these were further used to explore the metabolism of BDE-99 in vitro. The parent depletion approach was used for screening the ability of CYPs to eliminate BDE-99. Results: A reductively debrominated metabolite, BDE-47, and three oxidative metabolites, 2, 4, 5-tribromophenol, 5-OH-BDE-47, and 5'-OH-BDE-99, were identified from the BDE-99-treated rat hepatocytes, whereas no MeO metabolite was detected in the system. RT-PCR analysis showed that CYP 3A23/3A1, 1A2, and 2B1/2 were induced by BDE-99. Furthermore, using the heterological expressed CYP proteins in in vitro BDE-99 metabolism experiments we found that CYP1A2 and CYP3A4 showed the highest metabolic efficiency for BDE-99, with the metabolic clearance rates of CYP1A2 and CYP3A4 being 30.3% and 27.7%, respectively. CYP1A1 and CYP2A6 displayed relatively low clearance rates, while CYP2E1 seemed not to be associated with the BDE-99 metabolism. Conclusions: In our in vitro rat primary hepatocyte metabolism system, four metabolites of BDE-99 were identified, and CYP3A4 and CYP1A2 were demonstrated to be involved in the BDE-99 metabolism.

Abstract:With-the-flourishing-technological-development-of-college-and-the-expanding-college-run-industry,the-growing-problems-of-college-run-industry-limiting-the-further-development-have-been-so-noticeable.-By-analyzing-the-operation-and-management-of-college-run-industry,the-article-tried-to-explore-the-origin-of-the-problems-and-put-forward-the-corresponding-solutions.

Abstract:Objective:-To-make-a-full-understanding-of-medical-university-needy-students’-psychological-condition.-Methods:-SCL-90-self-rating-symptom-scale-and-self-made-“needy-students-mental-health-questionnaire”-were-applied-to-test-650-needy-students-of-a-medical-university.-Compared-the-results-with-some-regular-parametres,the-differences-among-various-groups-were-analyzed.-Results:The-number-of-needy-students-with-positive-criterion≥180-was-68,which-was-10.6%-of-the-total-number.-The-level-of-medical-university-needy-students’-psychological-condition-was-lower-than-local-adults,but-was-superior-to-local-university-students.-The-first-five-factors-of-the-needy-students’-psychological-problems-were-compulsion,personal-relationship,depression,psychoticism-and-hostility.-The-psychological-condition-of-the-needy-students-from-different-grade-was-remarkably-different.-The-needy-students’-psychological-condition-was-connected-with-their-grade,income,student-relations-and-their-academic-satisfaction.-Conclusion:The-general-psychological-condition-of-the-medical-university-needy-students’-is-comparatively-good.-But-there-are-some-distinguished-psychological-problems-among-the-minority-of-needy-students.-Educators-must-build-up-a-new-proper-psychological-education-system-according-to-the-various-characteristics-of-the-needy-students,such-as-grade-achievement-and-so-on,through-which-the-needy-students’-comprehensive-quality-might-be-effectively-improved.

Abstract:Objective: The cytochrome P450 17α-hydroxylase (CYP17) plays a vital role in androgen biosynthesis. A T-to-C polymorphism in the 5' promoter region of CYP17 has been implicated as a risk factor for prostate cancer, but the results of individual studies are inconclusive or controversial. To derive a more precise estimation of the relationship, we performed an updated meta-analysis from 31 studies based on 27 publications. Methods: A comprehensive search was conducted to examine all the eligible studies of CYP17 polymorphism and prostate cancer risk. We used odds ratios (ORs) with 95% confidence intervals (CIs) to assess the strength of the association. Results: Overall, individuals with CC/CT genotype were not associated with prostate cancer risk (CC vs. TT: OR = 1.03, 95% CI = 0.86-1.24, P = 0.72, Pheterogeneity < 0.0001; CT vs. TT: OR = 0.99, 95% CI = 0.87-1.12, P = 0.88, Pheterogeneity = 0.0006). In the stratified analysis by ethnicity, there was a significantly increased risk of prostate cancer among individuals of African descent under the recessive model (OR = 1.56, 95% CI = 1.01-2.39, P = 0.04, Pheterogeneity = 0.65). Conclusion: This meta-analysis suggested that CYP17 polymorphism might be associated with prostate cancer risk among individuals of African descent.

Abstract:Clinical-practice-is-an-important-part-of-the-medical-education-for-medical-students.-Based-on-clinical-practice,the-cultivation-of-legal-awareness-and-practical-ability-among-medical-students-in-clinical-practice-were-dicussed-in-this-article.-By-emphasizing-its-necessity-and-importance,the-author-analyzed-the-methods-to-cultivate-legal-awareness-and-practical-ability-among-medical-students-in-clinical-practice,and-then-put-forward-some-reasonable-proposals,which-was-of-great-significance-for-cultivating-high-quality-medical-graduates-and-building-a-harmonious-doctor-patient-relationship.

Abstract:It’s-an-important-task-for-clinical-educators-to-explore-effective-methods-of-humanistic-spirit-cultivation-for-medical-school-students.-The-authors-gained-valuable-experience-in-practice-on-the-humanistic-education-and-established-scientific-evaluation-system,which-should-integrate-humanistic-education-into-conventional-teaching-activities-and-develop-medical-students’-humanistic-spirit.

Abstract:How-to-guide-university-students-in-effective-leaning,-which-is-the-trend-of-education-of-leaning-society-and-life-long-leaning,-has-become-an-important-part-of-the-education-reform-in-all-the-countries-in-the-world-besides-China.-It-is-valuable-to-make-a-comprehensive-programme-of-leaning-guidance-for-university-students-through-comparing-with-other-counties-and-analyzing-the-problems-existing-in-our-county.

Abstract:Objective: CYP1A2 and NADPH-CYP450 oxidoreductase (POR) were expressed in the baculovirus/Spodoptera frugiperda (sf9) system. The aim of this study was to investigate the effects of heme precursors on the expression of CYP1A2 and POR. Methods: The heme precursors [δ-Aminolaevulinic Acid (5-ALA), Fe3+ and hemin] were introduced into the system to evaluate their effects on the expression of CYP1A2, POR and their co-expression. All the proteins were identified using immunoblotting, CO-difference spectroscopy, or cytochrome c assay. Results: In the present study, functional CYP1A2 and POR were successfully expressed in the baculovirus/sf9 system, and both of them showed high activities. Co-addition of 5-ALA and Fe3+ significantly improved expression of CYP1A2 by about 50% compared with the addition of 5-ALA, Fe3+ or hemin alone. Either co-addition of 5-ALA and Fe3+ or addition of 5-ALA or Fe3+ alone improved the POR expression level 2 fold and its activity 7-10 fold compared with control (no addition). However, unlike CYP1A2, there was no difference between the co-addition and addition of these heme precursors alone. Different ratios of BvCYP1A2 to BvPOR also affected the co-expression of CYP1A2 and POR, with a 3:1 ratio of BvCYP1A2 / BvPOR significantly increasing their co-expression. Surprisingly, the addition of 0.1 mM 5-ALA or Fe3+ alone, but not their co-addition, could significantly improve the CYP1A2 and POR co-expression (P < 0.05). Conclusion: 5-ALA and Fe3+ increased the expression of CYP1A2 and POR in a baculovirus/sf9 system, but the pattern of their expression was different between their expression alone and co-expression.

Abstract:Change-the-trend-of-social-changes-and-the-resulting-strong-social-oscillation,necessarily-through-social,school,family-and-other-sources-of-radiation-to-the-students-and-make-students-about-profound-changes-in-values.-The-survey-from-the-college-students-of-the-value-orientation-of-the-current-situation-as-the-starting-point,through-the-values-of-the-characteristics-of-our-students-and-causes-of-change,proposed-solutions-and-ways-to-achieve-that-I-set-the-correct-values-of-students,the-establishment-of-and-the-socialist-core-value-system-compatible-with-the-purpose-of-values.

Abstract:Objective: To explore the nature of deficiency in spleen-yin syndrome, which could provide scientific theoretical support and practical guidance for clinical Traditional Chinese Medicine (TCM) syndrome differentiation based on biology, and had a strong clinical significance. Methods: Serum Cu and Zn were detected by atomic absorption spectrophotometer, serum vitamin E by high performance liquid chromatography, serum vitamin C by 2,4-Dinitrophenylhydrazine Colorimetry, total superoxide dismutase (SOD) and Cu and Zn-SOD by the xanthine oxidase method, and malondialdehyde (MDA) by the 2-thiobarbituric acid method (TBA). Total antioxidant capacity was detected by a colorimetry kit. Amylase Activity was detected by an automatic biochemical analyzer. Lysozyme was detected by lysozyme detection plate, the diameter of bacteriolysis circle was measured and the corresponding content of lysozyme was obtained from a table of standard curve values. Results: No significant difference in total SOD and Cu, Zn-SOD was found between deficiency in spleen-yin group and normal group. However, such factors in deficiency in kidney-yin group were significantly lower than the other groups (P < 0.05). The MDA content in both deficiency in spleen-yin group and deficiency in kidney-yin group were significantly higher than that of normal group (P < 0.05), while the total antioxidant capacity was significantly lower than normal group (P < 0.05). The vitamin E content in deficiency in kidney-yin group was significantly lower than that in the other two groups (P < 0.05). No significant difference in the contents of vitamin C, Cu and Zn were observed in these groups. The Zn/Cu level in deficiency in kidney-yin group and the vitamin E level in deficiency in spleen-yin group decreased, but with no significant difference. Amylase activity in unit time in cases with deficiency in spleen-yin was lower than and had significant differences with that in normal cases, and higher than that in cases with deficiency in kidney-yin. The sectional velocity of saliva and the ratio of lysozyme in normal case group were significantly higher than other two groups, while deficiency in the spleen-yin group was significantly higher than the deficiency in kidney-yin group. Conclusion: All the results indicated that the objective pathological mechanism between the deficiency in spleen-yin and deficiency in kidney-yin was different.

Abstract:The-course“ideological-and-moral-cultivation-and-legal-basis”-for-case-teaching-has-the-ideological-and-political-orientation-on-its-content,the-different-levers-on-the-conclusion,and-complicated-selectivity-on-the-form,-which-including-that-teachers-select-cases-for-students-to-study,the-students-summarize-the-cases-for-teachers-to-comment.-The-case-selection-should-adhere-to-three-principles-on-ideology,direction-and-influence.-The-author-considered-that-the-leading-role-of-teachers,convincing-and-diversification-of-cases,initiation-and-creativity-of-students-were-the-major-factors-of-restriction-of-case-teaching,then-indicated-effective-methods-for-optimizing-case-teaching-such-as-reflecting-advanced-concepts-of“research-teaching”,creating-networks-platform,and-improving-the-comprehensive-evaluation-system.

Abstract:Through-a-year-of-reform-on-medical-courses-examination,the-authors-analyzed-the-characteristics-of-these-programs-which-focused-on-classroom-performance,clinical-performance,clinical-case-analysis-and-process-assessment,-summarized-the-effects-and-problems-of-the-reform,then-made-three-solutions,-which-were-teacher-training,teaching-reform-simultaneously-and-long-term-institutionalization.

Abstract:The-quality-oriented-education-is-a-kind-of-teaching-modes-improving-the-all-around-abilities-and-qualities.-In-the-course-of-anatomical-teaching-we-had-tried-our-best-to-exert-professional-function-into-the-medical-students,not-only-in-special-quality-and-ability,but-also-in-other-relative-qualities-and-abilities,which-was-beneficial-to-cultivate-the-talents-facing-the-modernization,the-world,and-the-future.

Abstract:Objective:To study the significance of human herpesvirus type 6 (HHV-6) infection on U251 astroglioma cells and its effects on biological functions of the cells. Methods:The model of HHV-6 infection in vitro was developed by infection of U251 cells with HHV-6. Synchronized U251 were infected with HHV-6. Cytopathic effect(CPE)were visualized by inverted phase contrast microscope. u22 gene of HHV-6 was amplified by PCR. The expression of immediate early protein 1 (IE1) and late protein(gB) were determined by indirect immunofluorescence assay(IFA). The proliferation of U251 cells was determined by MTT method,and the cell cycle was detected by flow cytometry analysis(FCM). Results:After 3 days,cells infected with the HHV-6 showed the typical cytopathic effect,cell swelling and the appearance of translucence and pleomorphism. u22 gene of HHV-6 was detected by PCR. HHV-6 immediate early protein 1 (IE1) and late protein (gB) were positive in the cultured cells by indirect immunofluorescence assay (IFA). HHV-6 could promote U251 cells proliferation by MTT assay. The FCM assay showed that the cycle of the infected U251 cells changed,compared infected groups with control groups,the former G1 phase cells decreased,and cells in S phase and G2-phase increased significantly. HHV-6 could promote U251 cells proliferation. Conclusion:HHV-6 can infect the U251 cells,cause cytopathic,stimulate cell proliferation,and change cell cycle. The results of this study indicated that HHV-6 might be correlated with the genesis and development of gliocytoma,but its mechanism is not clear.

Abstract:Objective:To construct the recombinant lentivirus containing HIV-1 Vif gene and detect the effect of Vif protein expression on the lytic cycle replication of KSHV. Methods:The fragment of Vif gene from expression vector pCI-neo-Vif was cloned into the lentivirus vector pHAGE-CMV-MCS-IzsGreen,then the recombinant plasmid pHAGE-Vif,packaging vector psPAX2 and envelope vector pMD2.G were cotransfected into the 293T cells. Culture media were harvested and filtered through a 0.45 μm filter to remove the cells. The viral titer was checked by observing the expression of green fluorescent protein(GFP). After infected with the recombinant lentivirus,the mRNA transcription and protein expression of Vif in 293T cells were detected by RT-PCR and Western blot,respectively. Then,BCBL-1 cells were infected by the recombinant lentivirus with the MOI of 1.0,and the protein expression of Vif,vIL-6 and Rta were detected by Western blot. Results:The recombinant lentivirus vector carrying Vif was constructed successfully with the viral titer of 4×107 efu/ml. BCBL-1 cells could be efficiently infected by the recombinant lentivirus with the MOI of 1.0,and Vif protein was readily expressed in these cells. Moreover,the expression of Rta and vIL-6 proteins of KSHV were significantly downregulated by Vif protein. Conclusion:Recombinant lentivirus with high titer and efficient infection of BCBL-1 cells could be obtained quickly and simply by using the lentivirus vectors system,and Vif expression could be observed in BCBL-1 cells infected with lentivirus-Vif. Furthermore,the Vif protein may inhibit the lytic cycle replication of KSHV. The results of this study lay the foundation for further studying on the molecular mechanisms and signaling pathways involved.

Abstract:Objective:To investigate the osteogeneous effect of recombinant retrovirus pLXSN/VEGF165 transfection on human marrow stroma cells(hMSCs). Methods:hMSCs isolated from health volunteers,cultured and purified in vitro were randomly divided into 4 groups. In retrovirus pLXSN/hVEGF165 group,recombinant retrovirus pLXSN/hVEGF165 was added into culture medium (1×1010 OPU/ml) and incubated for 24 h. In retrovirus pLXSN group,recombinant retrovirus pLXSN was added to take the place of retrovirus pLXSN/hVEGF165. In positive control group,dexamethasone (1 nmol/L),glycerophosphate (10 mmol/L) and ascorbic (50 mg/L) were added into culture medium for 2 weeks. In blank control group,no special treatment was administrated. Two weeks after treatment,VEGF expression were evaluated by immunohistochemistry,calcified nodes of hMSCs in each group were tested by Von Kossa stain and alkaline phosphatase (ALP) of culture medium in each group was detected. Results:hMSCs were uniform spindle-shape fibroblast after several passages. After treatment,hMSCs in retrovirus pLXSN/hVEGF165 group and positive control group gradually showed flat shape,but did not in retrovirus and blank control group. The level of VEGF expression in retrovirus pLXSN/hVEGF165 group were higher than the others,Von Kossa stain demonstrated that red bone collagen notes of hMSCs in retrovirus pLXSN/hVEGF165 group and positive control group were significantly more than that in retrovirus pLXSN and blank control groups. Similarly,ALP of culture medium in retrovirus pLXSN/hVEGF165 group and positive control group were significantly higher than those in retrovirus pLXSN group and blank control group. Conclusion:Recombinant of retrovirus pLXSN/hVEGF165 gene can transfect hMSCs and promote osteogenesis of hMSCs.

Abstract:Objective:It is proved that the pretreatment of oleanolic acid(OA) increases rat’s hepatic glycogen and alleviates the hepatic ischemic reperfusion injury(HIRI),but the mechanism is still not quite clear. This research is focused on the influence of OA pretreatment to rat’s PI3K-AKT-GSK-3β signaling pathway around its HIRI. Methods:Total of 128 male Sprague-Dawley(SD) rats were indvided into sham group(SH),ischemic reperfusion group(IR),0.5% sodium carboxymethycellulose(CMC-Na) group(CM),OA+0.5% CMC-Na group(OA) equally. Before operation all rats got intragastric administration one time a day for seven days,OA group with OA(100 mg/kg)+0.5% CMC-Na,SH group and IR group with water of the same volume,CM group with 0.5% CMC-Na of the same volume. At the 8th day,rats were suffered from segmental(70%) hepatic ischemia for 60 min and were followed by different periods of reperfusion. In the 8th day before the operation,the hepatic tissu were reperfused at the point of 0 h,3 h,and 6 h. The protein expressions of p-PI3K(p85),p-AKT(ser473),AKT,p-GSK-3β(ser9) and GSK-3β were evaluated. Results:Before operation,at the point of 0h,the protein expressions of p-PI3K(p85),p-AKT(ser473)and p-GSK-3β(ser9) in the OA group increased much more remarkably than other three groups(P < 0.05),while the protein expresions of SH group,IR group and CM group were no differences(P > 0.05). At the reperfusion point of 3 h,6 h,the protein expressions of p-PI3K(p85),p-AKT(ser473)and p-GSK-3β(ser9) in the OA group increased much more remarkably than other three groups(P < 0.05),while the protein expressions of SH group were lower than those of other three groups(P < 0.05). There was no difference between IR and CM group in the 3 proteins expression(P > 0.05). The protein expressions of AKT and GSK-3β in all the groups at all the points were no differences(P > 0.05). Conclusion:The pretreated OA increases the protein expressions of p-PI3K(p85),p-AKT(ser473)and p-GSK-3β(ser9) before and after HIRI,activating the signaling pathway of hepatic PI3K-AKT-GSK-3β,which may be one of the mechanisms of OA alleviating HIRI.

Abstract:Objective:To explore VEGF expression induced by PGE2 in human hepatocellular carcinoma cells HuH7 and the possible mechanisms of signal transduction pathways. Methods:HuH7 cells were treated with PGE2,EP1-4 receptor agonist,SQ22536+PGE2,H89+PGE2. VEGF mRNA and protein expression were examined by Real-time PCR,Western blot and ELISA in HuH7 cells. Results:The expression of VEGF mRNA in HuH7 cells were increased by 264%(P < 0.01),and the expression of VEGF in culture supernatant of HuH7 cells were increased by 24%(P < 0.01)after treated with PGE2(10 μmol/L) for 24 h. The expression of VEGF in cytoplasm of HuH7 cells were increased by 13.6%,25.7%,39.6% (P < 0.01)after treated with PGE2 (10 μmol/L) for 6 h,12 h,24 h respectively. The expression of VEGF in cytoplasm and culture supernatant of HuH7 cells were increased by 32.3% and 20.1%(P < 0.01)after treated with EP2 receptor agonist(10 μmol/L) for 24 h respectively,but have no changes after treated with EP1,EP3,EP4(10 μmol/L) receptor agonist. After the treatment of H89 (30 μmol/L) and SQ22536(500 μmol/L) for 24 h,the expression of VEGF in cytoplasm of HuH7 cells was decreased by 91.6% and 89.6%,and the expression of VEGF in culture supernatant of HuH7 cells was decreased by 98.3% and 91.9%,respectively,vs PGE2(P < 0.01). Conclusion:It is suggested that PGE2 might up-regulate the expression of VEGF through EP2 receptor of HuH7 cells,which could be partly related to the cAMP-PKA signal conduction pathway.

Abstract:Objective:To detect the expression of high mobility group protein A1(HMGA1) in HCC and investigate the relationship between the expression of HMGA1 and clinic pathological features of HCC. Methods:QRT-PCR was performed for detecting the expression of HMGA1 mRNA in 52 HCC tumor specimen and corresponding paracancerous liver tissues,10 normal human liver cancer tissue was used as negative control. The expression of HMGA1 protein in HCC was analyzed by immunohistochemistry. Results:The expression of HMGA1 mRNA was 6.82-fold higher in HCC tissues than in normal liver tissues (P < 0.05). And in HCC tissues with and without intrahepatic metastasis,the HMGA1 mRNA expression was 13.36 and 4.63 folds of that in normal liver tissues,respectively. The expression level of HMGA1 in HCC tissues was related to TNM stage,Edmonson grade and intraheptic metastasis(P < 0.05). The expression of HMGA1 protein was significantly higher in HCC with intrahepatic metastasis than in that without intrahepatic metastasis. Conclusion:HMGA1 expression was up-regulated in HCC,and it might be associated with the clinic stages and intrahepatic metastasis of HCC,and potentially used as an indicator for predicting the prognosis and a target for treatment approach.

Abstract:Objective:To investigate the clinical significance of Fascin expression in lung carcinoma. Methods:The clinical and pathological information of 63 cases of lung carcinoma were collected. The expression of Fascin in the tissues of lung carcinoma were detected and compared with that in paraneophastic tissues and normal lung tissues by Western blot. Results:The expression of Fascin in the tissues of lung carcinoma was higher than that in normal lung tissues(P < 0.05),which correlated with clinical stage(P < 0.05),lymphatic metastasis(P < 0.05) and histology grade(P < 0.05);There was significant difference between the expression of Fascin in squamous carcinoma and adenocarcinoma(P < 0.05);The expression of Fascin was not correlated with gender,age and the history of smoking(P > 0.05). Conclusion:The higher expression of Fascin in lung carcinoma is the marker of high invasive and low histology grade. Fascin may become a target in the therapy of lung carcinoma.

Abstract:Objective:To amplifiy of Trop-2 gene from clinic tumor tissue and express Trop-2 protein in E.coli. TOP10,and analyze its immunological characters with Western blot and ELISA. Methods:The Trop-2 gene was amplifiied by RT-PCR and cloned into pMD18-T. After sequenced,the Trop-2 gene was digested with NcoI and EcoRI and cloned into expression vector pBAD-gⅢ. The constructed vector had been identified by nucleotide sequence analysis,and then transformed into E.coli. TOP10. After induced with L-arabinase,the recombinant protein was purified with HisTrap affinity chromatography,and confirmed by Western blot and ELISA. Results:The Trop-2 was amplicated and sequenced,and the recombinant plasmid was constructed correctly. SDS-PAGE analysis showed that the recombinant protein was about 38 ku. The purified protein could be used as an antigen to detect the specific antibodies with Western blot and ELISA. Conclusion:The recombinant extracellular protein of Trop-2 could be expressed with high performance,and the antigenicity of the purified protein was certificated with the specific antibody.

Abstract:Objective:To investigate the effect of anti-CD3 plus anti-CD28 antibody co-stimulation on the release of interleukin-17 A (IL-17A) in human peripheral blood mononuclear cells (PBMCs). And the effect of triptolide on IL-17A release was also studied. Methods:PBMCs were stimulated with anti-CD3 monoclonal antibody (10 μg/ml) and anti-CD28 monoclonal antibody (10 μg/ml). ELISA was applied to measure the concentration of IL-17A in the supernatant of the cell culture. Results:IL-17A was released in time-dependant manner by PBMCs due to anti-CD3 plus anti-CD28 co-stimulation,however the release could be inhibited by triptolide at the concentration of either 100 nmol/L or 500 nmol/L. Conclusion:IL-17A produced by PBMCs might play an important role in the mechanism of asthma. Triptolide might inhibit the Th2 type allergic inflammation in asthma through reducing the release of the IL-17A.

Abstract:Objective:To explore the effects of IL-6 on the expression of Toll like receptor-4(TLR4) of THP-1. Methods:Flow Cytometry(FCM) was used to detect the expression of TLR4 of THP-1 cells,which was stimulated with different concertrations of IL-6(0 ng/L,10 ng/L,100 ng/L,1 μg/L,10 μg/L and 100 μg/L),and on different time after treated with 10 μg/L IL-6 (0 h,3 h,6 h,12 h,24 h,48 h). Results:Different concertrations of IL-6 can significantly promote the expression of TLR4 in THP-1 cells,especially under the concentration of 10 μg/L IL-6 group. Still this effect can be observed in 10 μg/L IL-6 group under treated for 3 h, 6 h,12 h,24 h and 48 h,especially at 24 h. Conclusion:The expression of of TLR4 on THP-1 cells can be promoted by IL-6 in a dose-and time-dependence.

Abstract:Objective:To investigate the preparation,extraction and identification of exosomes(Dex) derived from the different maturity dendritic cells(DCs) in vitro. Methods:F344 rat bone marrow-derived DCs were cultured 6 days then divided into 2 groups,the immature dendritic cells group(immature DC,imDC):the supernatant was collect;the mature dendritic cells group(mature DC,mDC):the concentration of 1μg/ml of LPS was added,and after cultured for 48 hours,the supernatant was collected. The Dex was isolated by multi-step centrifugation. Flow cytometry was used to detecte Dex phenotype. Enzyme-linked immunoassay (ELISA) was used to detected the IL-10 and interleukin-12 (IL-12p70) levels of in the supernatant. Both groups of Dex were co-cultured with imDC derived from Wistar bone marrow. The co-cultured DCs ability to stimulate T cell proliferation was detected by mixed lymphocyte reaction(MLR). Results:Compared with exosomes secreted from mDc,the imDC secreted exosomes expressing moderate levels of MHC-Ⅱ,low levels of CD80,CD86,CD40. And it has a weak stimulation to IL-12p70 secretion after co-cultured with Wistar rat bone marrow-derived imDC(P < 0.05),as to as stimulation to T lymphocyte proliferation(P < 0.05). Conclusion:The exosomes secreted by different maturity of DCs have different functions,may play different roles in the host immune response.

Abstract:Objective:To observe the effect of olfactory ensheathing cells (OECs) conditioned medium on NF,synapsin and synaptophysin expression in PC12 cells,and investigate the mechanism of olfactory ensheathing cell conditioned medium to promote growth of nerve cell differentiation. Methods:Primarily cultured OECs were prepared and purified,and the purified cell conditioned medium was collected into the olfactory ensheathing cell culture medium,then used to culture PC12 cells in 24 h,48 h,72 h. In vitro the morphological changes of the PC12 cells were observed under a phase contrast microscope;The expression and distribution of NF was determined by immunofluorescence methed;The relative content of NF,synapsin and synaptophysin proteins was detected by Western blot. Results:PC12 cells have long processes cultured in OECCM,and with the extension of the culture time,the cell morphology resembles neurons. Immunofluorescence staining showed the distribution of NF initially in the perinuclear. After the effects of OECCM the distribution of NF,increased and the ratio to β-tubulin was significantly high(P < 0.05).Western blotting showed the expression level of NF synapsin,and synaptophysin was significantly higher in OECCM group than in control group(P < 0.05). Conclusion:Olfactory ensheathing cells can secrete a number of cytokines to promote PC12 cell differentiation. These cytokines can promote the assembly of NF skeleton in PC12 cells,and induce neuronal function protein expression. So the cells can differentiate to the neurons in the structure and function.

Abstract:Objective:To compare improved‘ceiling’tissue culture method with digestion culture method to choose one method which can culture preadipocytes more properly. Methods:Cells obtained by these two methods,cultured 3 days,were determined by microscope for their self-differentiation. Preadipocytes proliferation and cell cycle were measured by MTT spectrophotometry and flow cytometer. After cultured 9 days,cells were counted to draw growth curves. After induced by DMEM/F12 with insulin,thyrine and transterrin(ITT) 5 days,the diameters of cells were determined by Image-Pro plus5.0 software and the content of lipid was measured by Oil Red O staining. Results:A few adipocytes were obtained by‘ceiling’tissue culture method appeared in the first generation cells. The results of proliferation and differentiation are similar in the cells obtained by the two methods. But ‘ceiling’tissue culture method can obtain more preadipocytes and need less money than digestion culture method. Conclusion:Comared with digestion culture method,improved‘ceiling’tissue culture method is a more simple and economic method which can obtained more preadipocytes

Abstract:Objective:To explore the ERRα expression in cardinal ligament and the effect of 17-β estradiol(17-βE2) on the proliferation of fibroblasts derived from cardinal ligament in patients with uterine prolapse. Methods:The tissues of cardinal ligament were collected from 10 patients with uterine prolapse and 10 normal controls for the detection of ERRα expression by flow cytometry;total of 10 cell strains of cardinal ligament fibroblasts derived from patients with uterine prolapse were established by primary cell culture and were treated with different concentrations of 17-βE2 (0 nmol/L,0.1 nmol/L,1.0 nmol/L,10.0 nmol/L,100.0 nmol/L) for 48 h. The effect of E2 on cell proliferation was then measured by the proliferous index(PI). Results:① ERRα were expressed in cardinal ligament of patients with uterine prolapse and normal controls,and the ERRα expression level of cardinal ligament was higher in normal controls than that in patients with uterine prolapse(P < 0.05). ② The PI of fibroblasts at 10.0 nmol/L 17-βE2 concentration was higher than that of control. The PI of fibroblasts at 100.0 nmol/L 17-βE2 concentration was lower than that of control. Conclusion:①The ERRα expression level of cardinal ligament decreased in patients with uterine prolapse. ② The effect of different concentrations of 17-βE2 on the proliferation of fibroblasts derived from cardinal ligament in patients with uterine prolapse was different:17-βE2 promoted fibroblasts proliferation at concentration of 10.0 nmol/L,but it induced apoptosis of the cells at 100.0 nmol/L concentration.

Abstract:Objective:To investigate the effect of oxidative stress on high glucose-induced apoptosis in neonatal rat cardiomyocytes. Methods:MTT assay was carried out to detect the viability of cultrured cardiomyocytes exposed to different concentrations of glucose ranging from 10 mmol/L to 50 mmol/L. Laser confocal microscopy,TUNEL assay and flow cytometry were designed to observe and detect cellular apoptosis. Cardiomyocytes were randomly divided into three groups:the control group,the high glucose group and the antioxidant group. Oxidative stress was evaluated by detecting levels of LDH,MDA and SOD in the supernatant of culture media. Results:High glucose was found to inhibit cardiomyocyte viability in a concentration-dependent manner. The results of laser confocal microscopy,TUNEL assay and flow cytometry all suggested that high glucose can induce cardiomyocyte apoptosis. Our findings showed significant increase of LDH and MDA levels and decrease of SOD activity in the high glucose group compared with the control group. However,in the antioxidant group,LDH and MDA levels decreased and SOD activity increased significantly compared with those in the high-glucose group. The apoptotic rate of cardiomyocytes in the high glucose group increased significantly compared with that in the control group,while the apoptotic rate in the antioxidant group declined remarkably in comparison with that in the high glucose group. Conclusion:Oxidative stress is an important mechanism for high glucose-induced cardiomyocyte apoptosis.

Abstract:Objective:To investigate the effects of soy isoflavone on the proliferation of primary cultured rat aortic endothelial cells (RAECs) induced by angiotensin Ⅱ. Methods:RAECs were cultured by tissue sticking method. Cell proliferation model was established by stimulation with AngⅡ. Cell proliferation was measured by MTT assay to observe the effects of various concentrations of soy isoflavone and NG-nitro-L-arginine methyl ester(L-NAME,100 μmol/L) on RAECs proliferation induced by AngⅡ. Nitric oxide (NO) level was measured by Griess reagent. Nitric oxide synthase (NOS) and inducible nitric oxide synthase (iNOS) levels were detected by chemical colorimetric method. mRNA expression of iNOS was measured by reverse transcription polymerase chain reaction (RT-PCR). Results:Soy isoflavone at the doses ranging from 0.1 μmol/L to 1.0 μmol/L inhibited cell proliferation in a dose and time-dependent manner. The inhibitory effects were partly blocked by 100 μmol/L of L-NAME. Soy isoflavone markedly increased NO,NOS and iNOS levels,and increased iNOS mRNA expression in RAECs. Conclusion:Soy isoflavone could inhibit RAECs proliferation induced by AngⅡ,and NO might contribute to this inhibitory effects.

Abstract:Objective:To explore the effects of rosmarinic acid (RA) on the tubular epithelial-mesenchymal transition by aldosterone (ALD) and its possible mechanisms in vitro. Methods:HK-2 cells were divided into five groups:normal group,HK-2 cells induced by ALD,ALD+RA (5 μg/ml),ALD+RA (25 μg/ml),ALD+rotenone(ROT,10 μmol/L). The morphology of transdifferentiate tubular cells was observed using phase-contrast-microscopy. E-cadherin,Vimentin,α-SMA and ERK1/2 were determined by semi-quantitative RT-PCR and Western blot;Reactive oxygen species(ROS) was detected by fluorescence microscopic and fluorescence elisa. Results:HK-2 cells induced by ALD converted into spindle shape from typical epithelium shape,the expression of Vimentin,α-SMA,phosphor-ERK1/2 and ROS production was significantly increased,the expression of E-cadherin was significantly decreased;compared with ALD treated groups,RA (5 μg/ml,25 μg/ml) and mitochondrial respiratory chain complex I inhibitor rotenone (ROT,10μmol/L) significantly inhibited ROS generation,vimentin,α-SMA and phosphor-ERK1/2 expression,and E-cadherin expression was restored. Conclusion:RA inhibited ALD-induced EMT via blocking mitochondrial-originated,ROS-dependent ERK1/2 activation.

Abstract:Objective:To identify and analyze the differential expression of L-lactate dehydrogenase B chain(LDH-B)in the penis of rat offspring after maternal exposure to di-n-butyl phthalate(DBP). Methods:Twenty pregnant rats were randomly divided into two groups and given DBP by gastric intubation at the dose of 800 mg/kg per day in the experimental group or 5 ml per day soybean oil in the control group from the 14 th to 18 th day of pregnancy. Penises were harvested from the fetal rats of the normal and exposed groups respectively at GD l9 .The expression of LDH-B detected and analyzed by Western blot and immunohistochemistry. Results:LDH-B expressed both in the experiment group and the control group,but the expression level is much lower in the experimental group than in the control group(P < 0.05). LDH-B was mainly located in the cytoplasm of the penis’s tissue. Conclusion:LDH-B protein expressed much lower in the penis of the rat offspring after maternal exposure to DBP than in the penis of the rat offspring without exposure to DBP.

Abstract:Objective:To compare the effects of different dosage of dexamethasone (DEX) on rat lung morphogenesis. Methods:Total of 120 newborn rats were divided into three groups randomly:a small dosage DEX group,a large dosage DEX group and a control group. On the 10th,14th,21st,45th days after birth,the histologic structures of 10 rat lungs in each group were observed with light microscope. Results:Under the light microscope,during the first 2 weeks,rat lungs in the two treatment groups had fewer alveolar numbers,larger alveolar space,and thinner alveolar septum compared with those in the control group(P < 0.01). On the 21st and 45th days,the airspaces of rat lungs in the two treatment groups were still enlarged,while the alveolar spetums were thickening. An“emphysematous”condition of the lungs could be observed especially in the large dosage DEX group. Conclusion:Using DEX postnatal could promote lung morphogenesis obviously,while with the dose increased,the normal alveolization was inhibited,and the impact continued well into adulthood.

Abstract:Objective:To observe and discuss the effects of Xuebijing injection on the expression of P-selectin in serum and on the pulmonary vascular endothelial cells of PTE model rats. Methods:The PTE model was established by intravenous injection of autologous blood clots. One hundred and five healthy male SD rats were randomly divided into control group,PTE model group,and Xuebijing-treated group. Every group were divided into 5 subgroups according to the time of 1 hour,3 hour,1 day,3 day and 7 day after opreation,with 7 rats in each subgroup. Pathological changes of lung tissue were observed,and the expression of P-selectin protein on the pulmonary vascular endothelial cells was evaluated by Western blot. The contents of sP-selectin in serum were detected by ELISA. Results:The notable embolism in pulmonary arteries and inflammatory cell infiltration around alveoli could be found by microscopy. The expression of P-selectin protein on the pulmonary vascular endothelial cell and the contents of P-selectin in serum significantly elevated from 1 hour after embolization (P < 0.01). Whereas after using Xuebijing injection,the pathological injury was ameliorated.The expression of Ps protein on the pulmonary vascular endothelial cells were significantly decreased from 1 hour after embolization (P < 0.01). The contents of P-selectin in serum were also decreased and the differences between PTE model group and Xuebijing-treated group were statistically significant from 1 day after embolization(P < 0.01 or P < 0.05). Conclusion:Xuebijing injection may reduce the damage of inflammation and coagulation-fibrinolysis imbalance to the lung tissue through decreasing the expression of P-selectin in PTE,so it has a certain protective effect to the lung tissue.

Abstract:Objective:To study the expression levels of thromboxane A2(TXA2) and prostacyclin (PGI2) in the plasma,endothelin-1(ET-1)in vascular endothelium,bronchial and alveolar epithelia of lungs,and hemodynamic changes in acute pulmonary thromboembolism(APTE) of rabbit,and to explore the effects of thrombolytic(urokinase,UK)on their expressions. Methods:Twenty four rabbits were randomly divided into a control group,a APTE model group,a UK therapy group,with 8 rabbits each. The APTE model was established by intravenous injection of autologous blood clots. The plasma’s concentrations of TXA2 and PGI2 were examined by radioimmunoassay before and 1 h,2 h,4 h,8 h after injection. The hemodynamic monitoring were used by float catheter. The expression levels of ET-1 in lung vascular endothelium,bronchial and alveolar epithelia were examined by immunohistochemistry. Results:①The radioimmunoassay showed that TXA2 and PGI2 had no diference in the control group at each point;TXA2 and PGI2 in the APTE group increased at 1 h,peaked at 2 h,decreased at 4 h,which also were higher than those in the control group at 1 h,2 h,4 h(P < 0.01). In the UK group,TXA2 and PGI2 increased at 1 h,peaked 2 h,decreased after 2 h,also decreased faster than those in the APTE group at 1 h,2 h and 4 h,respectively(P < 0.01). There were no differences in TXA2 and PGI2 before injection and 8 h after injectionin in each group(P > 0.05). ②Hemodynamic monitoring showed that the PAMP and CVP were no change in the control group;and PAMP and CVP in the APTE group and UK group increased immediately after embolism,peaked at 2 h,then decreased,which also were higher than those in the control group at 1 h,2 h,4 h,8 h(P < 0.01). In the UK group PAMP and CVP also decreased faster than those in the APTE group at 1 h,2 h respectively(P < 0.01). There were no differences at 4 h and 8 h between the APTE group and the UK group. ③The immunohistochemistry results showed in the APTE,the expression levels of ET-1 in lung vascular endothelium,bronchial and alveolar epithelia were significantly higher compared with those in the control group. The UK group was significantly lower in ET-1 as compared to the APTE group. Conclusion:After APTE,TXA2,PGI2,ET-1 participated increasing of pulmonary vascular resistance,and early thrombolytic treatment could decrease TXA2,PGI2,ET-1,also relieved pulmonary vascular resistance.

Abstract:Objective:To study the effect of lithium chloride pretreatment on spatial memory in aged rats’ hippocampuses after exploratory laparotomy. Methods:Twenty-four male SD rats,aged 18 month,were divided into three groups randomly (n=8 each):control group (C),operation group (O) and lithium chloride group (L/O). The rats in group L/O were injected with 2 mmol/kg lithium chloride into their peritoneal once a day and continued seven days,while the rats in the other two groups were injected with same volume of normal saline. All rats were established exploratory laparotomy model on eighth day. After 24 h,the rats’ spatial memory was measured by Morris test. Results:The latency of group C was lower than that of group L/O or group O(P < 0.05);the latency of group L/O was significantly lower than that of group O(P < 0.05);the swimming distance of group C was significantly shorter than that of group L/O or group O(P < 0.05);the swimming distance of group L/O was significantly shorter than that of group O(P < 0.05). Conclusion:Spatial memory was injured after exploratory laparotomy in aged rats. However,lithium chloride pretreatment could obviously improve these changes. It is suggested that lithium chloride pretreatment has potential effects for spatial memory decrease in aged rats after exploratory laparotomy.

Abstract:Objective:Parkinson’s disease (PD) animal model is an indispensable tool for exploring the etiology,pathogenesis,pathology,symptoms,signs,neuroprotect and drug therapy. Among so many animal models of PD,the most widely used currently are 6-hydroxydopamine (6-OHDA) lesioned rat model and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced mouse model. In order to clarify their own characteristics,it is compared and evaluated the preparation principles,production styles,measurement experiments,difficulties,stabilities and other consideration factors of the two kinds of animal models of PD in this article,with a view to make better choices and take full advantages of that in the research of PD.

Abstract:Objective:To investigate the association between MTHFR C677T polymorphism and toxicities of methotrexate-induced toxicities of childhood acute lymphoblastic leukemia. Methods:A total of 98 children with acute lymphoblastic leukemia were selected. And PCR-RFLP was used to determine the genotypes of MTHFR C677T polymorphism. The toxicity response of patients received high-dose methotrexate chemotherapy was observed. Results:Compared with the wild-type CC,the carries of the variant genotypes (CT and TT) had higher methotrexate plasma concentration. Conclusion:MTHFR C677T polymorphism is associated with the toxicities of methotrexate-induced toxicities of childhood acute lymphoblastic leukemia,mainly gastrointestinal reaction and bone marrow suppression.

Abstract:Objective:To investigate the mutation of non-syndromic sensorineural hearing loss (NSHL) and identify its molecular etiopathogenisis. Methods:Peripheral blood samples were obtained from 22 NSHL patients collected by out-patient clinic. Their genomic DNA were extracted from peripheral blood by extraction kits to undergo polymerase chain reaction and sequencing so as to detect the mutations of GJB2,SLC26A4 and mitochondrial 12SrRNA gene. Results:Of 22 patients,three mutations of GJB2 gene were found:2 have GJB2 235delC heterozygous mutation,2 have 235delC homozygous mutation and 1 have 176del16bp+299delAT double mutation;2 mutaions was found out in MtDNA 12SrRNA gene:C1494T and A1555G;SLC26A4 gene have 1 IVS7-2 homozygous and 2 heterozygous mutations. Conclusion:The PCR combined DNA sequencing is a classic and effective method in genetic diagnostic of hereditary deafness,able to identify the molecular etiopathogenisis of NSHL. But this method exist many shortcomings,such as unable to qualitate,time and money consuming. Furthermore,it can not detect multiple mutations of different genes at the same time. Therefore,a new method,which has advantages in genetic diagnosis of NSHL,such as low time and money consuming,high performance and accuracy,must to be developed. These advantages make it fit to be used in clinic gene testing of hereditary deafness.

Abstract:Objective:To study the gene mutation of thyroid hormone receptorβ(TRβ)in a family with thyroid hormone resistance syndrome accompanied by Hashimoto’s thyroiditis. Methods:The peripheral blood samples of the patient,9 family members and 9 healthy subjects were collected,then thyroid function was detected and the genomic DNA was isolated. The exons 1~10 of TRβ gene were amplified by PCR. The products of PCR were purified and sequenced directly to detect the gene mutation. Results:No mutation was found in exon 1~10 of TRβ gene. Conclusion:The mutation of TRβ gene is not only reason for the pathogenesis of thyroid hormone resistance syndrome.

Abstract:Objective:To explore the correlation of sex hormones concentration,ER receptor,and the polymorphism of ERβ gene with the development of gastric adenocarcinoma in young people. Methods:Two hundred and sixteen cases of primarily gastric adenocarcinoma in young people and 160 cases of gastrohelcoma surgery patients were analyzed. Radioimmunoassay(RIA) was used to determine the levels of sex hormone. Immunohistochemistry method was used to detect the expression of ER. Polymerase chain reaction-restriction fragment length polymorphism method was used to detect the polymorphism of ERβ of test group. Results:Serum androgenic hormone levels were significantly lower in the control group than that of the test group,but serum estradiol levels were higher in the control group than that of the test group. No significant difference can be seen in the TNM staging in the test group between male and female. But androgenic hormone levels decreased progressively,estradiol levels increased progressively with differentiation degree degraded,and there are significant difference between positive and negative lymph node metastasis. There was negative expression of ERα and ERβ in control group. The positive rate of ERα and ERβ in the test groups were 57.4%(124/216) and 15.7%(34/216). The positive rates of ER were significantly higher in the tissue with lower differentiation degree,having lymph node metastasis and deeper infiltration degree than that with higher differentiation degree,having no lymph node metastasis and shallower infiltration degree. Moreover,the positive rates of ER were higher in the stage of TNM Ⅲ than in the stage of TNMⅠandⅡ. RR and Rr genetype rate were higher than rr genetype rate. RR genetype was most obviously observed by PCR-RFLP with the allelomorphic gene distribution of RsaI. RR were significantly higher in the tissue with lower differentiation degree,having lymph node metastasis,deeper infiltration degree,and with the stage of TNM Ⅲ than that with higher differentiation degree,having no lymph node metastasis,shallower infiltration degree,and with stages ofⅠandⅡ. There was no significant difference in all genetypes with the allelomorphic gene distribution of Alu I in the test group. Conclusion:The alteration of sex hormones concentration in serum is one of important factors effecting the cancerization of gastric gland. The polymorphism of ERβ gene may be related to the development of gastric carcinoma.

Abstract:Objective:To evaluate the early prognostic role of plasma brain natriuretic peptide(BNP)and monocyte chemoattractant protein-1(MCP-1)levels in patients with unstable angina(UA). Methods:Eighty-five cases UA without heart failure were divided into IB(n=25),IIB(n=30)and IIIB(n=30)groups according to Braunwald classification, and 20 cases stable angina pectoris(SAP)were chosen in the control group. Plasma samples for BNP and MCP-1 were obtained in the first 24 h. The incidences of main abnormal cardiac event(MACE)were recorded during 30 days. Results:In every UA groups,plasma levels of BNP and MCP-1 elevated significantly,and in comparison to those in the control group,there were statistically significant difference(all P < 0.05). Plasma levels of BNP and MCP-1 in IIIB group were higher than those in IIB group and IB group(all P < 0.05). The levels of BNP and MCP-1 were significantly higher in patients who had complex lesions compared with those who had simple lesions(P < 0.05). Multivariable logistic regression analysis revealed that BNP and MCP-1 were all risk factors for cardiac events(all P < 0.05). Conclusion:Measurement of BNP and MCP-1 are helpful in predicting the short-term prognosis in UA patients without heart failure.

Abstract:Objective:To evaluate the effects of massive whole-lung lavage on pulmonary function and six-minute walk distance in patients with coal workers’ pneumoconiosis. Methods:Twenty-nine patients with coal workers’ pneumoconiosis were recruited. Pulmonary function tests and six-minute walk test were completed before and 2 weeks after massive whole-lung lavage respectively,and the symptoms were recorded. Results:Following massive whole-lung lavage,dyspnea and chest tightness were alleviated significantly,but no remarkable change was found in cough and sputum. Baseline pulmonary function testing identified 17 patients with damage of small airway or obstructive ventilation dysfunction. Compared with parameters before massive whole-lung lavage,maximal ventilatory volume (MVV) and six-minute walk distance were significantly improved [(96.8±14.1)% vs (91.2±15.2)%;(448.8±60.7)m vs (422.6±46.2)m],and total lung capacity (TLC) was significantly reduced [(119.4%±15.0)% vs (115.8±14.7)%,P < 0.05]. Conclusion:This study indicated that massive whole-lung lavage can improve symptoms,pulmonary function,exercise tolerance and life quality in patients with coal workers’ pneumoconiosis.

Abstract:Objective:To determine the levels of of MPO expression in human serum and placenta during pregnancy and their correlation with the development of preeclampsia. Methods:Patients with mild preeclampsia(n=33)and severe preeclampsia(n=30)were recruited while thirty-one women with normal pregnancies served as controls.Serum samples from uterine venous blood and peripheral venous blood and placentas were collected. Serum concentrations of MPO were determined with a chromatography method whereas the expression of MPO in placentas was detected by immunohistochemistry. Results:① Serum levels of MPO in uterine venous blood from both mild and severe preeclampsia groups were significantly higher than those of healthy controls(P < 0.01),with the highest levels detected in severe preeclampsia group(P < 0.01). ② Although serum levels of MPO in the peripheral venous blood from both mild and severe preeclampsia groups were higher than those of healthy pregnant group,there were no significant differences among three groups. ③ The levels of MPO expression in placentas of preeclampsia groups were significantly higher than those of healthy pregnant women,with the highest levels detected in severe preeclampsia patients(P < 0.01;P=0.017). Conclusion:The increased levels of MPO in uterine venous blood and placentas are closely correlated with the development of preeclampsia,suggesting that MPO may play a role in the pathogenesis of preeclampsia via local oxidative stress.