Abstract:The Confucian middle way is naive dialectics, which explores interdependency and associativity of the opposites in a contradiction. But the middle course excessively exaggerates the status and function of “neutralization”, negates and attempts to prevent the transformation of a contradiction, restrict its content, and falls to the verge of metaphysics. The metaphysical faultiness of the middle course reflects that the middle dialectics is not thoroughness.

Abstract:The ideology-related work at college is an essential constituent of the Party’s ideology-related work, the arena and forefront where various political parties compete for ideological dominance. Responding to the ideology-related events,the college should tightly hold on to the Party’s leadership and guidance of college ideology work on the basis of its adherence to the principles of seeking truths from facts, increasing compatibility and initializing predictions. Besides,the college should enhance the construction of the ideological arena,and the coordinated system of ideological education involving university,family and society. At the same time the college should also cultivate and improve college students’ abilities of self perfection,self motivational, self education and self management.

Abstract:In order to meet the demands of Chinese researchers of counseling and psychotherapy,answer for the needs of corresponding discipline construction,promote the process of theoretical study,as well as timely reflect the international trend on the development,exploring the schools of counseling and psychotherapy has become urgent. Researchers should keep a foothold on the schools of western psychology,as well as consider the theories and methods of counseling and psychotherapy as backbone,then considerate the internal logic of counseling and psychotherapy,construct a school conforming with China’s situation as well as servicing for counseling and psychotherapy,and inspire Chinese scholars’ awareness of innovation and initiation.

Abstract:The implementation of“the regulations of the disclosure of government information” meant the China’s information disclosure system was established through the form of laws and regulations formally. For the first time,it established people’s legitimate rights and interests of the government information disclosure in the scope of administrative litigation, and made clear of the citizens’ right to know and the judicial protection. Accordingly,how to properly resolve the information public administrative disputes became a critical task of people’s court. In this artide, based on the administrative litigation law, combined with “the regulations of the disclosure of government information ” and “on the trial of administrative cases about government information disclosure provisions of a number of issues”(that is“the openness of government information of judicial interpretati-On”),the eathor expressed some opinion of the court case-accepting scope,litigant,judicial and the problem of administrative compensation.

Abstract:The article detailedly described the subject,process and result of the negotiation of Australian pharmaceutical,then,summarizd the effect of inspiration to China, such as keeping independence of negotiations,paying attention to drug evaluation in economics and exploring new methods of price negotiation

Abstract:The effective utilization of scientific research resource is the key point to improve the level of medical undergraduates’ education. This article was based on the questionnaire survey from 412 undergraduates of Nanjing Medical University in two consecutive years, indicating their cognition and utilization status of scientific research resource. The survey described the cognition and using experience of scientific research resources’ connotation, infrastructure, personnel, funds, practice opportunities and information. The results indicated that the understanding of these resources and utilization were still in the primary stage and some key problems had not been solved satisfactorily with considerable space of promotion. Finally, the authors put forward countermeasures and prospects to work out the problem, then to improve the efficiency of research resource utilization in medical undergraduate education, enhance scientific research quality of medical students, and provide valuable experience for undergraduates’ education.

Abstract:Through the analysis of undergraduate research activities carried out in Nanjing Medical Univers-

Abstract:Innovative talents in the area of public health are required to the employment of prevent and control diseases. Renovating the idea of education and reforming the approach of talents culture are key points for developing innovative talents. Building the scientific research thinking and ability is the breekthrough point to develop the innovative ability of preventive medicine undergraduates through changing the teaching contents and methods,combining the scientific research and teaching,and establishing and optimizing the systems of operation,evaluation and encouragement.

Abstract:Mentoring plays an important role in foreign students’ clinical practice. Nanjing First Hospital affiliated to Nanjing Medical University,as a pilot,first introduced mentoring to foreign students’ clinical practice. The hospital initially established a new mentoring model for foreign students at clinical practice and then implemented it in accordance with PDCA cycle (plan-do-check-assessment) principle. After two years of operation,it achieved good practical results,which was welcomed both by students and teachers. However,there also existed some problems. In this article,the determination,implementation,supervision and evaluation (four PDCA cycle control) of mentoring model adopted by Nanjing Medical University have been analyzed to discuss its achievements as well as its problems,and then improvement measures were brought forth accordingly.

Abstract:Case discussion in general surgery tearching was taken for grade 2005 foreign interns in our hospital. Foreign interns took part in case discussion actively and had a positive evaluation effect. Compared with grade 2004,grade 2005 foreign interns had significantly improved in answering questions and achieving scores of rotation tests. Case discussion was an effective way to improve the level of clinical practice of foreign students.

Abstract:Based on the student-centered and learning-for-use teaching philosophy,the teaching reform had been conducted on the English course for master candidates with respect to curriculum development,teaching content,teaching and assessment method,to promote the students’ academic writing skills and oral communication competence. Certain achievements had been made and such advice as compiling appropriate medical English material and constructing online teaching platform were put forward to obtain the best teaching effect.

Abstract:Positive responces of readers’comments could effectively increase their satisfaction, which would be helpful for library to improve its service level and quality. In the current study,the questionnaire was used to investigate the students’ comments and suggestion for the construction and service lvevl of library,which were then sorted into three parts including collection resources,environment and facilities,management and service. Finally, the suitable proposals were provided for the improvement of library construction and service quality.

Abstract:To provide the suggestion for improving the hospital management,strengthening disciplines and training staff,statistical analysis of papers published from 2006 to 2010 of Jiangsu Cancer Hospital were analyzed from the following fields: the number of papers,the ratio of fund paper,grade of periodicals,title and educational background,published papers from graduate students in the key clinical disciplines. The results showed that the number of SCI was shown increasing trend,and the quality of published papers was significantly higher from doctors and seniors than those from others. It played the key role in increasing the paper output by means of strengthening disciplines and personnel training,more actively applying for research projects,increasing the awards, and establishing an effective evaluation system.

Abstract:According to the biologic immune system and the characteristics of the development of universities and colleges,the article explored three functions—“defense”,“constraints”,and“monitoring”of the internal audit, then put forward practical advice and suggestion,including establishing a suitable internal audit mode,supporting the sustained and healthy development of universities and colleges through using internal audit.

Abstract:Objective:To construct the recombinant luciferase reporter plasmids containing series of truncated sequences of Kaposi’s sarcoma-associated herpesvirus (KSHV) ORF50 promoter and seek the specific response sites sequences of ORF50 promoter acted by HSV-1. Methods:Series of truncated sequences of KSHV ORF50 promoter were amplified using PCR from KSHV genome. The PCR products were further cloned into pGL3 basic vector to construct the recombinant luciferase reporter plasmids. The recombinant plasmids were confirmed by restriction enzymes digestion and sequence analysis. After infected with HSV-1, Vero cells were transfected with the ORF50 promoter-driven luciferase constructs to induce luciferase gene expression. Relative luciferase activity unit (RLU) was calculated and compared. Results:Series of truncated sequences of KSHV ORF50 promoter were isolated and cloned successfully. The promoter-driven luciferase expression of pGL3-95, pGL3-46 and pGL3-17 were declined significantly in Vero cells infected with HSV-1 when compared with pGL3-1500, pGL3-750, pGL3-375 and pGL3-185. Conclusion:The recombinant luciferase reporter plasmids containing series of truncated sequences of KSHV ORF50 promoter were constructed successfully. The specific response sites sequences of ORF50 promoter acted by HSV-1 are between -185 bp and -95 bp.

Abstract:Objective:To assess the migration and differentiation capacity of β-galactosidase(β-gal) transgenic mouse derived amniotic mesenchymal stem cells (AMSCs) in recipient wild type mice after the transplantation subcutaneously. Methods:AMSCs were isolated from amnion membrane of the second-trimester β-gal transgenic mice and were cultured in either α-MEM containing 15% FBS, osteogenic- or adipogenic-induced medium. The second passage AMSCs derived from β-gal transgenic mice were transplanted subcutaneously into newborn C57BL/6 wild type mice. After 8 weeks,various organs from recipient mice were fixed and stained for Lac Z or embedded in paraffin. Then 5-μm sections were cut on a rotary microtome. The paraffin sections were dewax and observed under microscopy, and stained by immunofluorescence for β-gal,glial fibrillary acidic protein (GFAP) and myelin basic protein(MBP). Results:The second passage AMSCs derived from β-gal transgenic mice were positive for Lac Z staining and could differentiate into osteoblasts or adipocytes under cultures with the osteogenic- or adipogenic-induced medium. Followed subcutaneous injection of the second passage AMSCs derived from β-gal transgenic mice into wild type mice for 8 weeks,Lac Z positive cells were detected in thyroid,lung,spleen and brain of recipient wild type mice.The double positive cells for β-gal and GFAP, or β-gal and MBP, were detected in brain of recipient wild type mice. Conclusion:AMSCs transplanted through subcutaneously can survive at least 2 months,and migrate into multiorgans through circulation and differentiate into organ specific cells to participate in self-renewal of normal tissues. Our results indicate that AMSCs can be used as seed cells for stem cell therapy to treatment various diseases.

Abstract:Objective:To determine the effect of isopsoralen on the osteogensis and adipogenesis of bone marrow mesenchymal stem cells(BM-MSCs). Methods:Rat BM-MSCs were cultured in the absence or presence of isopsoralen for 14 days,and resulting cells were stained with methylene blue, Oil Red O and detected for alkaline phosphatase(ALP). RNA and protein were isolated from cells,and expression levels of osteogenesis and adipogenesis related genes were evaluated by RT-PCR and Western blot. Results:①The efficiency of CFU-f and ALP positive CFU-f formation by BM-MSCs was enhanced,and the expressions of osteogenesis related genes were up-regulated significantly in isopsoralen-treated cultures compared with control cultures. ②The adipocyte forming efficiency and the gene and protein expressions of peroxisome proliferator-activated receptor-γ(PPARγ) were inhibited significantly in isopsoralen-treated cultures compared with control cultures. Conclusion:Our results indicate that isopsoralen can stimulate osteogensis and inhibit adipogenesis of BM-MSCs,and the stimulating effect of isopsoralen is better than psoralen.

Abstract:Objective:To ivestigate the possible effects on normal bone marrow derived mesenchymal stem cells(MSCs) when co-cultured with myeloma cell line U266. Methods:Bone marrow MSCs from normal subjects were co-cultured with U266 by transwell. On the given time point, the levels of TNF-α in the medium, the mRNA expression levels of Dickkopf 1(DKK1) and hepatocyte growth factor(HGF) were mesured. Results:No evident morphologic and proliferative alterations could be observed in bone marrow derived MSCs after co-cultured with U266, and there was also no detectable increase in the level of TNF-α in superant of the co-culture system. However, the mRNA expression levels of DKK1 and HGF were upregulated in bone marrow MSCs after 5 days’ co-culture with U266. Conlusion:The “cross-talk” between bone marrow MSCs and U266 induced dysregulation of DKK1 and HGF, which might be involved in the pathogenesis of multiple myeloma.

Abstract:Objective:To determine the differentiation capability of RAW 264.7 cells following subcultures. Methods:The cells were treated with receptor activator of NF-κB ligand (RANKL). The 6th,9th,13th,18th and 24th passages were treated with or without RANKL for 6 days. Tartrate-resistant acid phosphatase(TRAP) staining was used to observe the formation of osteoclasts. Results: RAW 264.7 cells at different passages all have TRAP-positive cells. There were no obvious differences among 6th,9th,13th and 18th in positive rate(P > 0.05),and the positive rate of 24th passage were significantly lower than that of other passages. Conclusion: RANKL can induce RAW 264.7 cells to form osteoclast. If cultured for long time, the differentiation capability of this cell line was reduced.

Abstract:Objective:To prepare chitosan nanoparticles as gene carrier of multiepitope DNA vaccine,and explore the loading capability and its protection ability to plasmid. Methods:The chitosan nanoparticles were prepared with ionic cross-linking method. The encapsulation rate was determined by UV spectrophotometer. The binding ability and protective effect of plasmid pIRES-TH-FL were evaluated by agarose gel electrophoresis analysis. The size,morphology and distribution were observed by transmission electron microscope. The diameter and zeta potential were measured by zetasizer. The pIRES-TH-FL-chitosan nanoparticles were transfected into 293T cells in order to examine their expression. Results:The encapsulation efficiency of pIRES-TH-FL-chitosan nanoparticles was 99.28% by UV spectrophotometer. Agarose gel electrophoresis showed that chitosan nanoparticles can effectively combined with plasmid pIRES-TH-FL and protected it from nuclease degradation.The mean diameter of pIRES-TH-FL-chitosan nanoparticles was about 300 nm and the mean zeta potential was 32.4mV.Western blot assay result confirmed that the DNA-chitosan nanoparticles can be expressed in 293T cells. Conclusion:pIRES-TH-FL-chitosan nanoparticles of homogeneous diameter were prepared and successfully expressed in vitro.

Abstract:Objective:To generate recombinant adenoviral vector containing CRT-HBsAg fusion gene for developing a safe, effective and HBsAg-specific therapeutic vaccine. Methods:The fusion of CRT and HBsAg gene was constructed by using polymerase chain reaction(PCR), endonuclease digestion and ligation methods, and then the fusion gene was cloned into pENTR/D-TOPO transfer vector after the base pairs of DNA(CACC) sequence was added to the 5′ end. Adenoviral expression vector(Ad-CRT/HBsAg)containing CRT-HBsAg fusion gene was constructed by homologous recombinantion. The linearized DNA plasmid of the recombinant adenoviral vector was transfected into human embryo kidney(HEK 293A) cells to package and amplify recombinant adenovirus. The recombinant adenovirus titer was characterized by using the End-dilution assay. The expression of the CRT/HBsAg fusion protein in Ad-CRT/HBsAg transfected 293A cells was detected by Western blot. Results:The CRT-HBsAg fusion gene was characterized by using PCR, and sequencing result revealed that the length and sequence were accurate. The recombinant adenoviral vector, Ad-CRT/HBsAg, was generated successfully. The titer of Ad-CRT/HBsAg was characterized as 2.68×1011 pfu/ml. The CRT-HBsAg fusion protein was expressed by HEK 293A cells correctly. Conclusion:Recombinant replication-defective adenovirus expression vector containing CRT/HBsAg fusion gene was constructed successfully, and this study has provided an experimental basis for further research of HBV gene therapy.

Abstract:Objective:To investigate the potential influence of IL-29 on the expressions of protease activated receptor(PAR)-1,2,3,and 4 on the membrane and in the cytoplasm of P815 mast cells. Methods:After being challenged with various concentrations of IL-29 for 2 h,6 h and 16 h, the P815 mast cells were collected and treated for flow cytometry to detect the membranous and cytoplasmic PARs expression. Results:Compared with the corresponding control,expression of PAR-1 induced by IL-29 was significantly decreased either on membrane or in cytoplasm of P815 cells, and the difference between membranous and cytoplasmic PAR-1 expression induced by IL-29 was not significant. IL-29 did little effect on the expressions of PAR-2,3,4 either on the membrane or in the cytoplasm of P815 cells as well as the corresponding control,though the differences between membranous and cytoplasmic PAR-2,3 and 4 were significantly detected. Conclusion:IL-29 can down-regulate the membranous and cytoplasmic PAR-1 expressed by P815 mast cells but has little effect on the expression of PAR-2,3 and 4 either on membrane or in cytoplasm of P815 cells. Though significant differences between membranous and cytoplasmic PAR-2,3 and 4 are present, either membranous or cytoplasmic PARs expression is representative while flow cytometry can be used to detect IL-29 modulating PARs expressed in P815 mast cells.

Abstract:Objective: To study the effect of substance P on the expression of protease activated receptors(PARs) in peritoneal macrophages from normal and allergic mice. Methods:Allergic mice were induced by intraperitoneal injection of ovalbumin(OVA). Mice were divided into normal saline, 0.2 μg/ml, 2.0 μg/ml, 20.0 μg/ml and 200.0 μg/ml SP groups respectively, with 6 mice in each group. The expressions of PARs on the surface of peritoneal macrophages were then analyzed by flowcytometry. Results:PAR-1, PAR-2, PAR-3 and PAR-4 expressions on the surface of peritoneal macrophages were all significantly decreased in allergic mice compared with normal mice(P < 0.01). Substance P had no significant influence on expressions of PARs in macrophages(P > 0.05). Conclusion:The intraperitoneal injection of substance P had no effect on the expressions of PARs in macrophages from mouse peritoneum, but OVA treatment did.

Abstract:Objective:To investigate the regulatory role of TLR7/8 ligand R-848 on IgE production of murine purified B cells. Methods:CD19+B cells were purified with MACS from splenocytes of murine, and cultured with base medium, and medium with anti-CD40 plus IL-4 in the presence or absence of R-848. The concentration of IgE was determined with ELISA,and the activation of B cells was determined with FACS. Purified B cells were labeled with CFSE, and after culturing the cells were harvested and the division of cells was determined with FACS. Results:R-848 could inhibit IgE production of purified B cells stimulated with anti-CD40 plus IL-4 in a dose dependent manner, and enhance the activation of B cells. Moreover, R-848 could inhibit the division of B cells stimulated with anti-CD40 plus IL-4. Conclusion:The inhibitory role of R-848 on IgE production might be related with a reduction in B cell division.

Abstract:Objective:To produce a monoclonal antibody (mAb) against human trophoblast cell-surface antigen 2(Trop2) with hybridoma technology and identify its immune specificity. Methods:The spleen cells from BALB/c mice immunized with pancreatic cancer cell line BxPC3 cells were fused with SP2/0 cells to produce a monoclonal antibody against human Trop2. Further, several immunological methods such as ELISA,IF,IP,MALDI-TOF MS,FACS and IHC were used to characterize the immune specificity of the mAb. Results:The results demonstrated that the specific mouse anti-human Trop2 mAb was successfully produced and can be secreted continuously and steadily. Moreover, this mAb can effectively recognize the transmembrane glycoprotein Trop2 over-expressed in human cancer cells and tissues. Conclusion:We successfully obtained mouse anti-human Trop2 mAb which may be potentially utilized for some tumor targeted therapy.

Abstract:Objective: To make a fusion protein constructed of human anti-EGFR scFv/truncated protamine(scFv/tP) gene and analyze the binding activity of the over-expressed protein. Methods:Two pairs of oligonucleotide primers were designed and used to amplify the VH and VL nucleotide sequences,then use VH and VL as template to amplify the scFv. Then the protein gene was cloned into expression vector pBAD-A. The synthesized tP coding sequence was cloned into expression vector pBAD-AscFv,and expressed in E.coli TOP10F′. The fusion protein was detected by SDS-PAGE and Western Blot,then purified by NiNTA chelating agarose. The antigen binding activity was detected by ELISA. And the DNA binding ability was detected by gel shift assay. Results:Restriction digestion and DNA sequencing proved that scFv/tP was correctly cloned into expression vector. SDS-PAGE and Western Blot results showed that scFv/tP fusion protein was successfully expressed in TOP10F′. ELISA detection confirmed that the specific antigen binding activity of the fusion protein; and gel shift assay results suggested its DNA binding ability. Conclusion:The scFv/tP fusion protein successfully expressed in E. coli and could specially bind with both EGFR antigen and DNA.

Abstract:Objective:To investigate the effect of Resveratrol on oxidative stress and the expression of p27 protein in rat renal mesangial cells (RMCs) under high glucose. Methods:Cultured RMCs were divided into four groups:normal glucose (5.6 mmol/L), normal glucose plus Resveratrol(20 μmol/L), high glucose(30 mmol/L), high glucose plus Resveratrol at 20 μmol/L. After treatment for 72 h, the degree of RMCs hypertrophy was estimated through counting the ratio of the RMCs protein content to the cell number. The protein levels of p27 were detected by Western blot. The levels of malondialdehyde(MDA) in cell supernatant were assayed using chromatometry. The levels of reactive oxygen species(ROS) were detected by flow cytometry. Results:① As compared to the NC group, exposure of rat RMCs to high glucose at concentrations of 30mmol/L caused a significant increase in the levels of MDA and ROS(P < 0.01). The addition of Resveratrol attenuated high glucose- enhanced levels of MDA and ROS(P < 0.05).②As compared to the normal cells, exposure of rat RMCs to high glucose at concentrations of 30mmol/L caused RMCs hypertrophy and a significant increase in the protein levels of p27(P < 0.01). The addition of Resveratrol attenuated RMCs hypertrophy and high glucose-enhanced protein levels of p27(P < 0.05). while no notable difference at the protein levels of p27 was found between the Res and NC group(P > 0.05). Conclusion:Resveratrol may attenuate RMCs hypertrophy through reversing high glucose up-regulating oxidative stress and repressing the expression of p27 in RMCs.

Abstract:Objective:To screen differentially expressed genes in rat glomerular mesangial cells(GMCs)in response to sublytic C5b-9 by microarray of high flux. Methods:Rat primary GMCs were cultured and stimulated with or without sublytic C5b-9, then the total RNA in GMCs was extracted,and the RNA was reversely transcribed to biotinylated labeled cDNA probes. The mixed probes were hybridized for cDNA microarray. Then, the chips were subsequently scanned by Affymetrix Scanner 3000 and read with software GCOS1.4. Then differentially expressed genes were identified and annotated to Gene Ontology(GO)database in order to analyze these genes function. Results:The results showed that among expression profile of 31 000 genes, up-regulated and down-regulated genes over 2 folds under sublytic C5b-9 attacking for 40 min were 4 124 and 2 234, and were 4 512 and 2 859 under attacking for 3 h respectively. Meanwhile, there were 2 325 genes up-regulated and 1 419 genes down-regulated over 2 folds under sublytic C5b-9 attacking for 40 min or 3 h. Analysis of these differences of genes function by GO, these differentially expressed genes function mainly involved in regulation of biological process, biological regulation, localization,response to stimulus, developmental process,proliferation, metabolic process,cellular process, multicellular organismal process and so on. Conclusion:sublytic C5b-9 stimulating rat GMCs indeed could cause the changes of gene expression.

Abstract:Objective:To explore the effect of knockdown of Sig-1R on endoplasmic reticulum stress and podocyte injury. Methods: siRNA vectors for silencing Sig-1R were constructed and transfected into mouse podocyte clones(MPC5), which were divided into normal control, negative control and interference groups. The mRNA and protein expressions of Sig-1R were detected by real-time PCR and Western blot, respectively. The apoptosis of podocytes was detected by nuclear staining using Hochest 33258. The protein expressions of nephrin, desmin, bcl-2, bax, caspase-3, 8, 9, 12 were detected by Western blot. Results:The reducation of Sig-1R in mRNA and protein levels was detected when the podocytes were transfected with Sig-1R siRNA(P < 0.05). The expression of nephrin decreased, while the expression of desmin increased significantly(P < 0.05) compared to control. The apoptosis of podocyte was exhibited in the interference group. The expression of bcl-2 markedly decreased, while the expression of bax increased, and the ratio of bcl-2 to bax was less than 1/2.(P < 0.05). The expression of active caspase-3 and caspase-12 increased significantly(P < 0.05), while the expression of pro-caspase 9 decreased and pro caspase 8 had no change. Conclusion:The knockdown of Sig-1R can cause endoplasmic reticulum stress and podocyte injury.

Abstract:Objective:To investigate the effects of PCSK9 siRNA on CD36,SR-A1 and SR-B1 expressions in THP-1 derived macrophages. Methods:The siRNA for PCSK9 gene were transfected into THP-1 derived macrophages using positive ion liposome Lipofectamine 2000. Transfection efficiency was assessed by fluorescence microscope assay. The expression of PCSK9 in THP-1 derived macrophages at 24 h after transfection was detected by RT-PCR and Western blot. The most efficient siRNA was selected for transfection. At 24 h after transfection,macrophages were treated with ox-LDL for another 24 h. Then the intracellular lipid accumulation was observed by oil red O staining,and expressions of CD36 mRNA,SR-A1 mRNA and SR-B1 mRNA were detected by RT-PCR. Results:siRNA of 80 nmol/L showed the strongest inhibition for the gene expression of PCSK9,and ox-LDL-induced accumulation of cholesterol and upregulation of CD36 mRNA expression in THP-1 derived macrophages decreased by application of PCSK9 siRNA(P < 0.05). However,PCSK9 siRNA caused no effect on ox-LDL-induced upregulation of SR-A1 mRNA and SR-B1 mRNA expression in macrophages. Conclusion:PCSK9 may be involved in atherosclerosis development by inhibiting the expression of CD36.

Abstract:Objective:To investigate the effect of CRP on the [Ca2+]i in mice aorta endothelial cells(ECs). Methods:Mice aortic endothelial cells were isolated and cultured. ECs were divided into control group and CRP treated group,and the changes of [Ca2+]i was examined after 25 min. For the effect of atorvastatin,the cells were pre-incubated 30 min by atorvastatin(1 μmol/L)prior to adding CRP(100 mg/L). Results:[Ca2+]i of ECs significantly increased after the treatment of CRP in a dose dependent manner. Furthermore,atorvastatin partially abrogated the up-regulation of [Ca2+]i. Conclusion:Inflammatory of ECs was up-regulation in response to CRP stimulation. Meanwhile,the research also demonstrated that atorvastatin partial suppressed inflammatory induced by CRP treatment.

Abstract:Objective: To explore the role of MAPKs signal transduction system in the proliferation of human mammary carcinoma cell line MCF-7 induced by insulin. Methods:Human mammary carcinoma cell line MCF-7 was treated with different concentrations of insulin(0 nmol/L,25 nmol/L,50 nmol/L,100 nmol/L,200 nmol/L). MTT assay was used to explore the proliferation of MCF-7 cells with or without MAPKs signals blocked by their specific inhibitors which were SP600125, the inhibitor of the phosphorylation of JNK,and PD98059, the inhibitor of the phosphorylation of ERK1/2. The expressions of phospho-JNK and phospho-ERK1/2 at different time treated with insulin were detected by Western blot, and changes of those proteins expressions were observed when MAPKs signals blocker was added. Results:Insulin can promote the proliferation of MCF-7 cells in a concentration-dependent manner. Western blot result implicated that 100 nmol/L insulin induced the activation of JNK and ERK. The phosphrylation of JNK was detected beginning at 5 minutes after insulin treated and sustaining for an hour, while the activation of ERK was at the 30th minute after insulin treated and later than that of JNK. Then we reevaluated the proliferation effect promoted by insulin after MAPKs signaling pathway blocked by their inhibitors. The results showed that after the activities of JNK, ERK and JAK/STAT inhibited, the proliferation effect of insulin was also attenuated. Otherwise,Western blot result showed that, while blocking the MAPKs signaling pathway,10 μmol/L PD98059 could inhibit the activation of ERK1/2 induced by 200 nmol/L insulin,while 20 μmol/L SP600125 and 20 μmol/L AG490 couldn’t inhibit the activation of ERK1/2 obviously. Conclusion: Insulin can stimulate the proliferation of MCF-7 cells in vitro, and MAPKs signal transduction system involves in the proliferation of MCF-7 cells, which could mediate and promote mammary carcinoma development.

Abstract:Objective:To explore the mechanism of pioglitazone(PGZ)on brown adipose tissue(BAT)function in insulin resistance and type 2 diabetes mellitus, by using high fat diet-induced obesity(HFD)mouse. Methods:The C57BL/6J mice were randomly divided into the normal diet group and high-fat diet(HFD)group. After 20 weeks,the HFD group mice were randomly divided into obese control group and PGZ treatment group.They were orally administered placebo and PGZ[10 mg/(kg·d)]respectively for one month. The levels of body weight,serum insulin and oral glucose tolerance were measured by Biochemistry technology; The effect of pioglitazone on brown adipose tissue(BAT)was assessed by real-time quantitative PCR and Western-blot. Results:The bodyweight, serum insulin levels increased(P < 0.05)and oral glucose tolerance decreased in HFD group. Comparing to obese controls,serum insulin,glucose levels significantly decreased(P < 0.05),and oral glucose tolerance increased in PGZ treatment group; The expressions of proteins and mRNA relative to BAT function increased in PGZ treatment group. Conclusion:PGZ can enhance the function of BAT by regulating BAT relative gene and protein expressions, and may be the important reason of improving insulin resistance and metabolism in HFD mouse.

Abstract:Objective:To comfirm the reliability for the neonatal rats model of periventricular leukomalacia caused by lipopolysaccharide. Methods:Sixteen mature pregnant wistar rats were divided into experimental(n=8)and control groups(n=8).On 15 days of gestation, LPS(0.3 mg/kg)and distilled water were injected intraperitoneally to experimental(n=8)group and control group, respectively. Seventy neonatal rats of experimental group and eighty-two in the control group were obtained till the gestation day of 21. Morphological changes and water content in brain tissues were studied. Results:①Periventricular leukomalacia,band-like and spotty coagulation necrosis were found in brain of neonatal rats in the experimental group, but not in the controls; ②Oligodendrocytes, mitochondrial obvious swelling , ridge decreased or vacuolization and endoplasmic reticulum expansion were observed in brain of neonatal rats, in the experimental group, but not in the controls;③Water content in neonatal rats' brain of the experimental group[(89.92±4.06)%]was significantly higher than that in the control group[(87.78±5.21)%](P < 0.05). Conclusion:Lipopolysaccharide given to maternal rats could result in periventricular leukomalacia and increase the water content in the brain tissues of neonatal rats.

Abstract:Objective:To investigate the influence of oleanolic acid(OA) pretreatment to rat’s IKK/I-κB/NF-κB signaling pathway around its hepatic ischemic/reperfusion injury(HIRI). Methods:One hundred and twenty eight male Sprague-Dawley (SD) rats were randomly divided into sham group(SH),ischemic/reperfusion group(IR),0.5% sodium carboxymethycellulose(CMC-Na) group(CM) and OA+0.5% CMC-Na group(OA). Before the operation,the rats of each group received intragastric administration of corresponding solution once a day for seven days(OA group with 100 mg/kg of OA dissolved in 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 then followed with different periods of reperfusion. The hepatic tissue were obtained at 0 h,3 h,and 6 h after the reperfusion. The expression of IKK2 and I-κBα in the cytoplasm and NF-κB p65 in the nucleus were evaluated by the method of Western blotting. Results:There was no IKK2 expression in the cytoplasm,and only few of NF-κB p65 was detected in the nucleus before operation or 0h after reperfusion in each group. Meanwhile,the volume of I-κBα detected in the cytoplasm has no significant difference among four groups at the same point of time. The expression of IKK2 in the cytoplasm and NF-κB p65 in the nucleus in the SH group was much lower than those in other three groups(P < 0.05),while those proteins detected in OA group is less than those in CM group and IR group(P < 0.05)at 3h or 6 h after reperfusion. At these two points of time,the expression of I-κBα in SH group was higher than that in the other three groups(P < 0.05),and this protein detected in OA group is more than that in CM group and IR group(P < 0.05).The expressions of IKK2 and I-κBα in the cytoplasm and NF-κB p65 in nucleus between CM group and IR group had no significant difference at each point of time(P > 0.05). Conclusion:HIRI would increase the expression of IKK2 in the cytoplasm,leading to the phosphorylation of I-κB,activating NF-κB and making it to translocate into the nucleus,and then the activated NF-κB which could mediate inflammatory response eventually caused damage of liver cells. The pretreatment of OA can inhibit the activating of the signaling pathway of IKK/I-κB/NF-κB,which may be one of the mechanisms for OA alleviating HIRI.

Abstract:Objective:To detect the effects of 17β-estradiol on the survival of cortical hippocampal neurons and determine whether it is associated with nNOS/PSD95 complexes. Methods:ICR mice was adopted to culture cortical hippocampal neurons. Eight days later, lactate dehydrogenase(LDH) release rate were examined in cultured cortical hippocampal neurons exposed to DMSO or 10 μmol/L 17β-estradiol for 30 min. The neurons were photographed through a light microscope. Meanwhile, nNOS/PSD95 complexes of neurons treated with DMSO,10 nmol/L 17β-estradiol and 10 μmol/L 17β-estradiol were measured by co-immunoprecipitation. Results:At 30 minutes after administration, 10 μmol/L 17β-estradiol treatment increased LDH release rate and nNOS/PSD95 conjugating in cortical hippocampal neurons, and 10 nmol/L 17β-estradiol treatment had no influence on nNOS/PSD95 conjugating. Conclusion:17β-estradiol of 10 μmol/L could injure cortical hippocampal neurons by promoting the conjugating of nNOS and PSD95.

Abstract:Objective:To investigated the blocking effect of high frequency alternating current signal to nerve action potential conduction. Methods:Frog sciatic nerve was dissected out in the experiments. The amplitude of action potential(AP) and the state of muscle during high-frequency sinusoidal stimulation were observed to determine the blocking effect. Results:The results showed that in a certain frequency complete block could be achieved at the threshold. When the stimulation amplitude increased upon threshold, there was no obvious difference in the character of the block. With an increase in frequency, block threshold also increased. The muscle tetanus due to the block frequency stimulation vanished after a short period signals whose frequency was higher than the block frequency were induced. In the following course, the muscle contraction was blocked by high frequency AC signal. Conclusion:Blocking effect of high frequency AC signal to nerve action potential conduction can potentially provide new methods for clinical applications of electrical nerve block in local anesthesia, acesodyne and spasmolysis.

Abstract:Objective:To research the effect of Bmi-1 gene on glioma cells apoptosis. Methods:Bmi-1 gene in U251 glioma cells is silenced by siRNA, and the effect was detected by RT-PCR, the apoptosis condition was detected by flow cytometry, and one way ANOVA and t test were used for statistics analysis. Results:Gel electrophoresis strip of Bmi-1 mRNA in RNA interfered U251 cells is weakly positive, those of negative controls and non-disposals are strongly positive. The gray scale ratio is statistically analyzed, and the difference between RNA interfered cells and non-interfered cells was significant. RNA Interfering may significantly down-regulate the expression of Bmi-1 in U251 cells The apoptosis rate of(32.53±6.33)% in interfered U251 cells is higher than that of(21.91±5.63)% in controls, and the difference is significant statistically. Conclusion:The Bmi-1 gene expression could inhibit the apoptosis of glioma cells and promote the genesis and development of glioma.

Abstract:Objective:To establish a new approach to evaluate cellular enzyme activity of glucosylceramide synthase(GCS) accurately by fluorescent thin-layer chromatography and spectrophometer,and decide its role in drug-resistant cancer cells. Methods:Using fluorescent NBD C6-ceramide,GlcCer/Cer and ceramide glycosylation of drug-resistant breast cancer cell(MCF-7-AdrR)was measured under different concentrations of NBD C6-Ceramide and increasing treatment time. Radioenzymatic assay were performed on these cells as control method. Protein expressions of GCS and P-gp were assessed by Western-blot analysis and immuno-fluorescence staining. Results:These data indicated that 5 μmol/L of NBD-C6-Ceramide was the optimal concentration,and GCS activity increased with time delay. This method has been used to accurately evaluate the levels of GCS enzyme in drug-resistant cells,which is consistent with radioenzymatic assay. Ceramide glycosylation in MCF-7-adrR and MCF-7-adrR/GCS cells was dramatically increased compared with that in MCF-7 cells(P < 0.01),while the level of glycosylation in MCF-7-AdrR/asGCS was much lower than MCF-7-adrR(P < 0.01). Conclusion:The cell-based fluorescent method is direct,reproducible for assessing ceramide glycosylation. It was applicable to validate GCS activity in drug-resistant cancer cells and would further help to measure GCS activity in vivo.

Abstract:Objective:To explore the molecular genetic characterization of two families with aminoglycoside-induced and nonsyndromic sensorineural hearing loss. Methods:Blood samples were obtained from 7 maternal members and 1 married-in spouse of the two families. Genomic DNA was extracted with conventional method. Firstly, 9 hot spots for mutations in four most common pathologic genes, GJB2, GJB3, SLC26A4 and mitochondrial 12S rRNA, were screened with the DNA microarray to detect the deafness-associated mutations. The whole mitochondrial genomes and nuclear modifier genes TRMU and MTO1 of two probands were then PCR amplified and submitted for sequence analysis. Results:Mitochondrial 12S rRNA C1494T mutation was detected in all 7 maternal members of the two families. Sequence analysis of the complete mitochondrial genomes in two probands revealed the distinct sets of mtDNA polymorphism (52 other nucleotide changes), in addition to the identical 12S rRNA C1494T mutation. None of these 52 variants, however, were shown to be pathogenic. The whole mitochondrial genome of proband from each of the two families was established that they belong to mitochondrial haplogroups D4 and D5a respectively. No mutations were identified in either TRMU gene or MTO1 gene. Conclusion:C1494T mutation in the mitochondrial 12S rRNA gene is the main molecular mechanism responsible for the hearing loss in the two pedigrees, and the use of aminoglycoside antibiotics may enhance the phenotypic manifestation of deafness-associated mitochondrial mutation. Mitochondrial haplogroups and nuclear genes (TRMU and MTO1), however, seems not play a role in the phenotypic expression of C1494T mutation in these two families.

Abstract:Objective:To investigate the application of transcatheter closure in patients with patent ductus arteriosus(PDA) with severe pulmonary arterial hypertension(SPH), and assess the medium and long-term results. Methods:Thirty seven patients with PDA underwent attempted transcatheter closure. On the basis of haemodynamic and clinical data obtained before and after attempted occlusion, the final PDA was carried out. The follow-up time was 1 week, 1,3, 6,12 months and every year after transcatheter closure. Results:The occlusion therapy was performed in 27 PDA. Systemic artery oxygen saturation before and after closure was (94.3±1.9)% and (96.2±1.4)%, respectively(P < 0.05). Systolic pulmonary arterial pressure (SPAP) decreased significantly from(90.1±17.2)mmHg to (45.1±10.9)mmHg (P < 0.05) and systolic arterial pressure increased significantly from (120.1±20.5)mmHg to (129.3±24.8)mmHg after closure(P < 0.05). The interventional occlusion was not carried out in 10 patients because of significant reduction of systemic artery oxygen saturation after six-minute walk test and no significant reduction of SPAP during the trial closure of PDA. In 27 patients with closure of PDA, SPAP was reduced to normal level in follow-up. Conclusion:Transcatheter closure is an effective treatment for patients with PDA associated with reversible SPH during the medium and long-term follow-up.

Abstract:Objective:To investigate the effect and mechanism of hydrocortisone on circulating T lymphocyte apoptosis in human septic shock. Methods:Total 57 patients with septic shock from January 2006 to January 2009 in Intensive Care Unit of Nanjing First Hospital Affiliated to NJMU were prospectively randomized into treatment groups and control group. Low-dose hydrocortisone was used in treatment group but not in control group. Peripheral venous blood samples of the included patients were obtained to determine the circulating T lymphocyte apoptosis by Annexin V and flow cytometry. At the meantime,the plasma concentration of TNF-α,IL-1β and IL-10 were determined by EASIA. All these results were compared with 20 healthy volunteers and 18 pyemia patients. In the meantime,the difference of septic shock control group and hydrocortisone treatment group was analyzed to investigate the effect of low-dose hydrocortisone on these results. Results:In baseline,the apoptosis percentage of CD4+T lymphocytes in healthy volunteers,pyemia patients and septic shock patients was (4.41±1.45)%,(7.87±3.82)% and (11.01±4.52)%,respectively. And the percentage of Annexin V-positive CD4+ T lymphocytes was higher in septic shock patients than in pyemia patients(P < 0.05),and the later was higher than healthy volunteers(P < 0.05). At the same time, there was no difference in the apoptosis percentage of CD8+ T lymphocytes among these groups(P > 0.05). After treatment of 48 h,the apoptosis percentage of CD4+ T lymphocytes was higher in hydrocortisone treatment group than that in control group,(P < 0.05),while,the apoptosis percentage of CD8+ T lymphocytes had no difference between the two groups(P > 0.05). The plasma concentration of TNF-α in healthy volunteers,pyemia patients and and septic shock patients in baseline was (16.44±9.55)pg/mL,(29.58±13.6)pg/mL and (47.99±25.63) pg/mL respectively. And it was higher in septic shock patients than in pyemia patients(P < 0.05). And the later was higher than healthy volunteers(P < 0.05). After treatment of 48 hours,the plasma concentradtion of TNF-α in hydrocortisone treatment group was lower than that in control group(P < 0.05). The plasma concentration of IL-1β in healthy volunteers,pyemia patients and septic shock patients in baseline was (13.12±5.07)pg/mL,(25.21±14.7)pg/mL and (22.94±22.01)pg/mL respectively. It was higher in pyemia patients and septic shock patients than that in healthy volunteers(P < 0.05). However,there was no difference between the pyemia patients and septic shock patients.(P > 0.05). After treatment of 48 hours,Threr was no difference in the plasma concentration of IL-1β between control group and hydrocortisone treatment group(P > 0.05). The plasma concentration of IL-10 in healthy volunteers,pyemia patients and and septic shock in baseline was(6.38±7.5) pg/mL,(9.67±4.88)pg/mL and(15.01±5.36) pg/mL respectively. It was higher in septic shock than in pyemia patients (P < 0.05). And the later was higher than healthy volunteers(P < 0.05). Forty-eight hours after onset,the plasma concentration of IL-10 in hydrocortisone treatment group was higher than that in control group(P < 0.05). Conclusion:Low-dose hydrocortisone can induce CD4+ T lymphocytes apoptosis when administered in septic shock,and it can also up-regulate the plasma concentration of IL-10. We concluded that IL-10 may be one of the reasons that low-dose hydrocortisone induced circulating CD4+ T lymphocytes apoptosis in septic shock.

Abstract:Objective:To compare the prognostic accuracy among three gastric cancer lymph node staging systems(pN,MLR and LODDS). Methods:Total 1 247 gastric cancer patients who received radical resection with complete follow-up data were included. ROC curves of Three Lymph Node Staging Systems were plotted and the area under curve(AUC)were statistically analyzed by grouped t test. Results:In the overall sample, samples with T2 or T3 stage,and samples with <15 or ≥ 15 nodes retrieved,all the three staging systems had prognostic value,but no statistical difference was found among AUCs of those three staging systems. Only LODDS had prognostic value in the sample without node metastasis and sample with T1 stage. However,in the sample with lymph node metastasis,all three had prognostic value,and the AUC differences between MLR and pN,LODDS and pN were statistically significant. Conclusion:The prognosic accuracy of MLR and LODDS is better than pN; MLR and pN are not able to predict prognosis of early gastric cancer;LODDS is the most accurate lymph node staging system for predicting the prognosis of gastric cancer.

Abstract:Objective: To compare the efficacy of ondansetron on the prevention of postoperative nausea and vomiting(PONV) under different general anesthesia methods. Methods:Two hundreds and forty patients undergoing gynecologic laparoscopy with different maintain methods of general anesthesia were randomly divided into six groups:sevoflurane group(group A1), sevoflurane with ondansetron group(group A2), propofol group(group B1), propofol with ondansetron group(group B2), propofol combined sevoflurane group(group C1), and propofol combined sevoflurane with ondansetron group(group C2), 40 patients in each group. Ondansetron of 8mg was given intravenous at 5 min before the end of surgery. Observed the nausea and vomiting situation after tracheal extubation in different periods(0~4h, 4~8h, 8~12h, 12~16h, 16~24h). Results:① In group B1 and group C1, the incidences of nausea reduced significantly compared with those in group A1 in 24 h(P < 0.05). In group A2, B2 and C2 compared with those in group A1, B1 and C1 respectively, the incidence of nausea was significantly lower in 0-12 h(P < 0.05), while within 12-24 h the incidence of nausea did not reduced obviously. ② In group B1 and group C1, the incidences of vomiting reduced significantly compared with those in group A1 in 24 h(P < 0.05). Compared with those in group A1, B1 and C1 respectively, the incidence of vomiting in group A2, B2 and C2 was significantly lower in 24 h(P < 0.05). Conclusion:Compared with the anesthesia methods of propofol or propofol combined sevoflurane, the sevoflurane anesthesia method has a more susceptible to PONV in patients. Ondansetron doesn’t have the same efficacy in PONV under different general anesthesia methods, and the treatment of nausea was better than vomiting within 24h after surgery. What’s more, ondansetron has better treatment of PONV caused by sevoflurane anesthesia method.

Abstract:Objective:To compare the value of diffused optical tomography(DOT)and color doppler flow imaging(CDFI) in the differential diagnosis of breast occupied lesions. Methods:Seventy-nine patients with eighty-five breast lesions were examined by DOT and CDFI. Using surgical pathology as the diagnostic standard, the accuracy of DOT and CDFI were compared in the diagnosis of benign and malignant breast masses. Results:The sensitivity,specificity and accuracy of DOT were 92.1%,87.2% and 89.4%,respectively. The sensitivity,specificity and accuracy of CDFI were 73.6%,76.6% and 75.3%,respectively. The sensitivity,specificity and accuracy of pulsed wave doppler u1trasonography(PW)according to blood flow resistance index(RI)were 57.9%,85.1% and 72.9% respectively. Combined with PW,the sensitivity,specificity and accuracy of CDFI were 55.2%,89.4% and 71.7%,respectively. Conclusion:The accuracy of DOT is higher than that of CDFI, thus it’s a new technique worthy of popularizing.