Abstract:Early embryos in vitro are special substance and have the highest level in legal objectivity system, because they have all genes developing to humans. Embryo in vitro produced by artificial insemination, the couples who had accepted the artificial reproductive technology had the ownership; Embryo in vitro produced by medical research, the medical research institutions had the ownership. The exercise of ownership of the owner should be restricted to “human dignity” and no person can sale or buy early embryos in vitro.

Abstract:Strengthening the education and management of floating Party members is the important component of grass-roots Party organizations construction. Based on the characteristics of floating Party members in Nanjing Medical University, modern information technology and innovative education management mode, the authors created a new online home of learning and teaching for floating Party members to exchange ideas.

Abstract:Objective: To evaluate health-related quality of life (HRQoL)of married women in childbearing age in rural China and explore the main factors affecting their HRQoL in order to make relevant policies and recommendations. Methods:EQ-5D measure and Visual Analogue Scale (VAS) were used to assess HRQoL. Results:The overall health of married women in childbearing age was better than the general population. Gynecological diseases were women’s main health problem. HRQoL of those with higher level education and without gynecological diseases was better than others. Conclusion:Married women in childbearing age should increase their awareness of health and deal with the pressure from family and work. The Government should carry out the census and treatment on women’s diseases periodically and pay more attention to women’s education. Relevant policies should be made to improve women’s health.

Abstract:Health is the common goal that human beings have constantly pursued. New features such as large pupulation, fast evolution and large differences presented as new characteristics of China national health as the “soft power” of a country is the device of social harmony and stability, the driving force behind economic development, a passport for international exchanges and cooperation, and the new requirment to fulfill international responsibility. At present, the country’s national health standards should be enhanced based on a national level in order to move towards a “health state”.

Abstract:Objective: To evaluate health-related quality of life (HRQoL) of rural residents in China and analyze health-related factors. Methods: EQ-5D instrument was used to measure the health state. Results: People’s health in east, middle and west China were different obviously. Women’s health was worse than men. People with higher education had better health while low-income population were meeting more health problems. More people had problems at the dimensions of “pain/discomfort” and “anxiety/depression”. Conclusions: Health policy making should pay more attention to people’s main health problems. Health of vulnerable groups such as women, the elderly, low-income people should be protected as a top priority in order to improve equity in health.

Abstract:The article analyzed employment status and pressure on undergraduate of the Health Service Management, and professional employment retated factors of students of the Health Service Management, discussed how to establish the professional image of the brand discipline and teaching reform, to ensure the quality of undergraduates education,successful employment and other related issues.

Abstract:In the process of constructing the harmonious society, the medical students＇ humanistic quality education is very important to establish harmonious doctor-patient relationship, promote the development of medical and health services, and advance the social harmony. In view of existing deficiencies in humanistic quality education of our medical colleges, the author puts forward some corresponding countermeasures, so as to realize the combination of medicine and humanities, and then contribute to a harmonious society.

Abstract:University lecture is an important content of science and culture construction for the campus and also an important support of humanity and science education for the students. However, in process of arranging lectures, there are obvious deficiencies in the lecture schedule, guidance, brand and characteristic, which hinder the function of university lectures. In view of existing situation, it is significant to innovate the construction of university lecture and promote the function of lecture in campus science and culture development in order to achieve higher education talent cultivation.

Abstract:In view of the characteristics of financial assistance for needy students in Taiwan in combin ation with the existing situation and problem of mainland financial assistance for needy students, the author suggested that the aiding systems, grant amount and supporting ideas should be improved.

Abstract:With the deepening education reform advanced by fast development of information technology, it is clear that the application of morden educational technology into clinical practice teaching has been a natural trend. On the basis of digital assisted instruction platform, Nanjing Drum Tower Hospital developes the curribulum of clinical practice and takes an effective management based on curriculam and information as well. As a result, the efficiency and outcomes of clinical internships are improved significantly.

Abstract:Objective: To probe into the feeling of the male nursing undergraduate students while learning fundamental nursing practical skills, to explore their study attitudes and influence factors, then provide corresponding measures. Methods:Sixteen male nursing undergraduate students of grade 2 were interviewed with the method of in-depth interview of qualitative research. The whole interviewing process was recorded. The data was analyzed, arranged and summarized for the themes. Results:Two themes were identified and descried that one was the attitude of learning fundamental nursing practical skills, such as good understanding of the importance but poor enthusiasm in learning, the another was the influential factors, including lack of professional sense,sex conflict, lack of belonging, heavy mental stress and fatigue. Conclusion:It is widely considered by the male nursing students that fundamental nursing practical skills is an important component of the nursing education. The present status of study attitudes towards leaning fundamental nursing technology is not ideal due to the lack of the needs of belonging, respect and self-esteem, self-actualization and so on.

Abstract:Directed by competency-based education theory,the article explored the application of the modules of employable skill(MES) in clinical teaching of prosthodontics. The clinic process was divided into differential successive teaching modules. Through several teaching methods, such as personal training, team-based learning and instructor assistance, MES was carried out to cultivate the high-quality graduates of stomatology.

Abstract:Objective: To investigate the effect of application of case-based learning in the course of ophthalmology teaching. Methods: The content of ophthalmology was divided into two parts, one part was experimental group which taught by case-based learning, and the other part was control group by traditional instructing. Then, a contrast of teaching effect was made between two groups. Results: There was no statistical significance in theory examination score while there were statistical significances in case analysis score and evaluation of teaching effect between two groups. Conclusion: Case-based learning has more advantages than traditional instruction, specifically on improving quality of students.

Abstract:Basic surgery is a bridge curriculum from foundational medicine to clinical medicine. The experimental teaching of it includes basic skill and synthetic surgery. The authors combined PBL and LBL teaching method and made a preliminary analysis of the teaching effects through objective-structured clinical examination in seven-years undergraduate Grade 2007. There were significant differences between combining PBL and LBL teaching method and traditional LBL teaching method, PBL teaching method in the aspects of test results, classroom atmospheres, teachers’ enthusiasm and students’ activity. The combination of PBL and LBL teaching method was proved to be effective for improving teaching quality.

Abstract:Da Yi Jing Cheng (the doctor with kindheartedness and justices) is the second introduction of Beiji Qianjin Yao Fang (Essential Recipes for Emergent Use Worth A Thousand Gold) wrote by Sun Simiao in Chinese Tang dynasty. With comprehensive contents, the introduction tells the doctor to promote the morality and medical skills. It is very important for the medical humanity education.

Abstract:Objective:To construct the recombinant secretory plasmid containing vIL-6 gene of KSHV,detect its protein expression in EA.hy926 cells,and observe the effect of the secretory protein on proliferation and angiogenesis of vascular endothelial cells. Methods:A pair of PCR primers for vIL-6 gene including HindⅢ and XhoⅠ restriction sites was designed according to the sequence of pvIL-6 F. vIL-6 gene was amplified by PCR, taking pvIL-6 F as template. Then purified vIL-6 gene fragments were digested and cloned into pSecTag2B vector to generate recombinant secretory expression plasmid named as pSecTag2B-vIL-6. pSecTag2B-vIL-6 was transfected into EA.hy926 cells. The expression of vIL-6 protein in EA.hy926 cells was detected by Western blot, and supernatant of 293T cells transfected by the recombinant secretory plasmid was collected, then the effect of vIL-6 on proliferation and angiogenesis of vascular endothelial cells by autocrine and paracrine was detected by cell proliferation assay and microtube formation assay. Results:The recombinant secretory plasmid carrying KSHV vIL-6 gene was constructed successfully. After transfected with the above plasmid, the exact band of vIL-6 was detectable by Western blot. EA.hy926 cells treated either with supernatants collected from 293T cells transfected by recombinant secretory plasmid or with transient transfection of the recombinant secretory plasmid, could perform an enhancement in proliferation and angiogenesis. Conclusion:The recombinant secretory plasmid pSecTag2B-vIL-6 was constructed successfully, and vIL-6 protein could be correctly expressed in EA.hy926 cells. Importantly, vIL-6 protein could promote the proliferation and angiogenesis of vascular endothelial cells through autocrine and paracrine pathway.

Abstract:Objective: To explore the method of cryopreservating human induced pluripotent. Methods:Colonies of induced pluripotent stem cells(iPS cells) were dissected into pieces(containing approximately 100~200 cells) using mechanical methods, then sequentially treated with 10%,20% vitrification solution, and sealed in mini straws with the second solution. Mini straws were then dropped into liquid nitrogen immediately for vitrification and cryopreservation. After vitrificated preservation, iPS cells were thawed in 37℃ water bath, and were immediately balanced in 0.2 mol/L sucrose solution and then in 0.1 mol/L sucrose solution, and at last plated on feeder layer cells. Alkaline phosphatase activity, expressions of OCT4, SOX2 in iPS cells and cell differentiation were evaluated. Results:Post vitrification and thawing, iPS cells maintain properties of pluripotent stem cells, including normal morphology, alkaline phosphatase staining,OCT4 and SOX2 expression. Calculation of colony recovery rates indicates that approximate(77.40±13.12)% of iPS cell colonies survived the freezing and thaw procedures. Conclusion:Vitrification with ministraws is a very useful and effective cryopreservation method for iPS cells.

Abstract:Objective:To establish C57BL/6J mouse embryonic stem (ES) cell lines with bFGF-free knockout serum replacement (KSR). Methods:C57BL/6J mouse blastocysts 3.5 days post coition(dpc) were collected and cultured in the medium supplemented with KSR and 1 000 U/mL LIF, with or without 10 ng/mL bFGF. The ES cell lines were identificated by morphology, alkaline phosphatase activity, karyotype, cell surface maker SSEA-1 and pluripotent markers (Oct4, Sox2 and Nanog) analysis. The differentiation ability of ES cells was analyzed both in vitro (embryoid bodies) and in vivo (teratoma). Results:Four cell lines were established out of 6 blastocysts cultured in the medium without bFGF; in contrast, no ES-like colony was observed when bFGF was added to the medium after isolation of 3 high quality blastocysts. The ES cells proliferated well without evidence of differentiation. The ES cells had typical ES-like morphology, expressed alkaline phosphatase,SSEA-1, pluripotent makers such as Oct4, Sox2 and Nanog. They could form embryoid bodies when been cultured in suspension and generate teratomas when been injected subcutaneously into SCID mice, consistent with the the pluripotent abilities of ES cells. Conclusion:KSR containing medium without bFGF could be an efficient culture system for the establishment of mouse ES cell lines.

Abstract:Objective:To investigate the effects of JAK/STAT6 pathway in airway remodeling in the mice with bronchial asthma and influence of AG490 on airway remodeling and its mechanism. Methods:Thirty-two femal BALB/c mice were divided into four groups at random. They were control group,the asthma group,the group of AG490 intervention and the group of budesonide intervention. The mice were sensitized and challenged with ovalbumin to establish the chronical asthma model. The airway inflammation and the alteration of airway structure were observed by the means of haematoxylin-eosin(HE) and MASSON staining. The expression of p-STAT6 was examined by Western blot and immunohistochemistry analysis,while the content of cyclinD1 was detected by Western blot and RF PCR. Results:HE and MASSON’s trichrome staining showed that there were bronchial smooth muscle hypertrophy, submucosa incrassation, airway stenosis, collagen fiber increase and mass inflammatiory cells infiltration in asthmatic group. In AG490 and budesonide intervention groups,the symptoms mentioned above were much more ameliorated. The Western blot analysis showed that the expression levels of p-STAT6 and cyclin D1 in asthmatic group were higher than control group(P < 0.01), but were decreased in AG490 and budesonide intervention groups(P < 0.01). The result of RT-PCR showed that the level of cyclinD1 mRNA in asthmatic group was higher than control group(P < 0.01), and was reduced in the AG490 and budesonide intervention groups(P < 0.01). Compared with the control group, the immunohistochemistry result showed that the expression of p-STAT6 was much higher in asthmatic group(P < 0.01), and between them were AG490 and budesonide intervention groups(P < 0.05). Conclusions:JAK/STAT6 signal pathway involved in airway inflammation and asthma remodeling. AG490 may ameliorate the progression of airway inflammation and remodeling via regulation of JAK/STAT6 signal pathway.

Abstract:Objective:To investigate effects of budesonide on the expression of phosphorylated vascular endothelial growth factor receptor 2(VEGFR2) in asthmatic mice with airway remodeling. Methods:Twenty-four BALB/c mice were randomly divided into control group, asthma group and budesonide intervention group. The mice were sensitized and challenged with ovalbumin(OVA) to establish the chronic asthmatic model. Budesonide (10 ml) was atomized half an hour before sensitization in budesonide intervention group. After finished the last sensitization for 24h, the mice were sacrificed. Right lungs were isolated and stained with haematoxylin and eosin (HE). Immunohistochemistry was used to detect the expression of phosphorylated VEGFR2, and Western blot was applied to examine the expressions of phosphorylated VEGFR2 and Erk in the left lungs. Results:There were bronchial airway remodeling and mass inflammatory cells infiltration in asthmatic group; budesonide intervention ameliorated the symptoms mentioned above significantly. The protein expression levels of phosphorylated VEGFR2 and Erk in asthmatic group were higher than those of the control group(P < 0.01), and both of them decreased dramatically in the budesonide intevention group(P < 0.01). Conclusion:The expressions levels of phosphorylated VEGFR2 and Erk in asthmatic mice were much higher than control group, and budesonide could inhibit phosphorylation of them, thus relieve airway inflammation and delay the process of airway remodeling.

Abstract:Objective: To investigate the effect of hypoxia-induced factor 1α (HIF-1α) gene silence on the expression of vasculogenic mimicry associated genes in human esophageal squamous cancer. Methods:Eca109 cells were divided into normal control group(non-transfection group),negative control group(transfected with empty plasmid) and experimental group(stably transfected with recombinant plasmid pGCsi-shHIF-1α), and were transplanted into nude mice to establish esophageal squamous cancer-bearing nude mice. The growth of transplanted tumor in nude mice were observed. HE staining was conducted to detect atypia and necrosis of tumors in each group. Immunohistochemistry and Western blot were used to observe the protein expression of HIF-1α and epithelial kinase (EPHA2), vascular endothelial cadherin(VE-cadherin), matrix metalloproteinase-2 (MMP-2) in tumor tissue. RT-PCR was adopted to detect the mRNA expression of HIF-1α,EPHA2,VE-cadherin and MMP-2. Results:The tumor formation and growth in experimental group were lower than the control groups. The volume and weight were decreased significantly than the control groups(P < 0.05). HE staining confirmed the significantly decreased atypia and increased necrosis in experimental group. Immunohistochemistry and Western blot results showed that HIF-1α protein expression of the experimental group was silenced, and EPHA2, VE-cadherin expression were decreased significantly (P < 0.05). RT-PCR results suggested that the mRNA levels of HIF-1α and EPHA2, VE-cadherin, MMP2 in experimental group were decreased to a different extent, EPHA2 and VE-cadherin were significantly reduced(P < 0.05). Conclusion:HIF-1α gene silence can suppress the growth of esophageal squamous cancer in vivo and inhibit the VE-cadherin and EPHA2 expression in the formation of vasculogenic mimicry,HIF-1α may participate in the regulation of vasculogenic mimicry through these two sites in esophageal squamous cancer.

Abstract:Objective:To evaluate the feasibility of preparation of reprogrammed neural stem cells derived from skin and neural differentiation in vitro. Methods:Skin-drived precursors (SKPs) were isolated and purified from rat skin tissue,identified and seeded at passage 3 in 12-well culture plates. SKPs were divided into three groups:group A was transfected with neurogenin2 via GFP-plentivirus;group B was transfected with GFP-plentivirus;group C was cultured without transfection. All groups were induced by common inducer for a week after 7 days of transfection.The morphology of induced SKPs was observed by microscope,and the markers of neural cells(MAP-2 and NeuN) were detected by immunocytochemistry. Results:The green fluorescence of SKPs in group A reached the peak on the 7th day after transfection and existed until the 8th passage.The shapes of SKPs were fusiform or ellipse, and most SKPs express several apophyses after inducement. Immunofluorescence demonstrated that 90.98% of the SKPs expressed MAP-2 and 62.23% of the SKPs expressed NeuN. SKPs in group B expressed green fluorescence and had no significant typical morphological changes after inducement. Immunofluorescence demonstrated that 34.35% of the SKPs epressed Map-2 and no SKPs expressed NeuN;SKPs in group C did not express green fluorescence and had no significant typical morphological changes after inducement. Immunofluorescence demonstrated that 29.54% of the SKPs expressed MAP-2 and no SKPs expressed NeuN. Conclusion:The reprogrammed neural stem cells can be prepared through transfecting neurogenin2 into SKPs via plentivirus in vitro, and their capability of differentiating into neural cells elevated visibly after neurogenin 2 transfection.

Abstract:Objective: To construct the eukaryotic expression plasmid carried mouse S100A16 gene short hairpin RNA(shRNA), and to investigate the silencing effect of shRNA on the S100A16 gene in mouse 3T3-L1 cells. Methods:siRNAs were designed according to the S100A16 cDNA sequence in GenBank,and inserted into plasmid PLKO.1-sP6-GFP. The recombinant plasmids were identified by PCR and sequence analysis, and transfected into 3T3-L1 cells with LipofectamineTM 2000. The transfection efficiency was reported by green fluorescence protein expression and gene inhibition efficiency was analyzed by realtime RT-PCR. Results:The correct sequences of recombinant PLKO.1-S100A16-GFP-shRNAs were confirmed by DNA sequencing. Green fluorescence observed by fluorescence microscope showed that shRNAs had been transfected into 3T3-L1 cells(transfection rate was 90%). And the expression of S100A16 mRNA was effectively down-regulated by PLKO.1-S100A16-GFP-shRNA2(inhibition rate was above 70%). Conclusion:The eukaryotic expression plasmids of S100A16 shRNAs were constructed successfully. The PLKO.1-S100A16-GFP-shRNA2 could down-regulated the expression of S100A16 gene effectively. This is fundamental for the study on the function of S100A16 gene.

Abstract:Objective:To study the effect of transfecting the neuronal guidance factor Netrin-1 and enhanced green fluorescent protein gene(EGFP) to rat bone mesenchymal stem cells(BMSCs) by adenovirus on the cytokine secretion, differentiation and tube formation. Methods:BMSCs were separated and purified in vitro. Surface receptors of adenovirus infected BMSCs and cytokines were detected by real-time PCR or RT-PCR.Tube formation of HUVEC was observed by coculture with BMSCs. BMSCs migration was observed by Transwell experiment. Results:Adenovirus infected BMSCs expressed some kinds of Netrin-1 receptors and secreted vascular endothelial growth factor(VEGF),basic fibroblast growth factor(bFGF),brain derived neurophic factor(BDNF) and nerve growth factor(NGF). BMSCs cultured with Netrin-1 and VEGF markedly increased the tube formation of HUVEC and BMSCs migration,compared with Netrin-1 or VEGF alone(P < 0.05). Conclusion:BMSCs successfully infected with Ad5-Netrin-1-CMV-EGFP can express EGFP and Netrin-1.Netrin-1 has the ability to accelerate tube formation and BMSCs migration and VEGF can enhance the effect when used together.

Abstract:Objective:To investigate the correlation between polymorphism of Lys198Asn of endothelin(ET)-1 gene exon 5 and ventricular septal defects(VSD) related pulmonary arterial hypertension(PAH),and risk factors of PAH. Methods:Genotypes were determined by the PCR-restriction fragment length polymorphism(PCR-RFLP) method in 140 VSD related PAH cases and 140 non-PAH controls. Results:①The frequencies of GT and TT genotypes in PAH group were significantly higher than non-PAH group (43.6% vs 33.6%, adjusted OR=1.78,P < 0.05;10.7% vs 3.6%,adjusted OR=3.34,P < 0.01; respectively);②The frequency of GT+TT genotypes in PAH group was significantly higher than that of the controls(54.3% vs 37.2%, adjusted OR=1.94,P < 0.01);③The T allele frequency in PAH group was significantly higher than that in the controls(32.5% vs 20.4%, P < 0.01). Conclusion: ET-1 gene Lys198Asn polymorphism is associated with and may be a new risk factor of PAH.

Abstract:objective:to investigate the relationship between tumor necrosis factor-β(tnf-β) rs1041981 gene polymorphisms and coronary artery disease(cad) in nanjing city. methods:a case-control study comprising 399 patients with cad and 280 controls was performed. single nucleotide polymorphisms of rs1041981 in tnf-β gene were genotyped and analyzed using the high-flux taqman probe technology coupled with fluorescent quantitative pcr method. results:the frequencies of the cc, ca, and aa genotypes of rs1041981 were 25.1%, 50.3% and 24.6% in cad group and were 41.1%,41.8% and 17.1% in control group respectively. there were statistically significant differences in the distributions of the genotypes (χ2=20.1,p < 0.05). the risk of cad in those of ca and aa genotypes was 1.38 times of cc genotype(or=1.38,95%ci:0.84~2.05). the serum tnf-β and high sensitive c-reactive protein levels of the cad group were significantly higher than that of the control group(p < 0.05).conclusion:the single nucleotide polymorphism (snp) of rs1041981 in tnf-β gene is associated with cad in nanjing area and the allele a may be a risk factor of cad.

Abstract:objective:to study the expression,distribution and biological significance of ribonucleotide reductase m2 subunit(rrm2) in human cholangiocarcinoma tissues and qbc939 cell line. methods:the expression of rrm2 was evaluated immunohistochemically in 40 cholangiocarcinoma samples,40 cases of adjacent histologically noncancerous bile duct,and 10 specimens of normal bile duct tissue. the expression of rrm2 in human cholangiocarcinoma cell line qbc939 and human normal bile duct epithelial cell line hibec were also detected by rt-pcr and western blot analysis. results:rrm2 was expressed in 72.5%(29 of 40) samples of carcinoma tissues,but it was undetectable in all adjacent histologically noncancerous bile and normal bile duct tissue. rrm2 was also expressed in cholangiocarcinoma cell line qbc939 but not in hibec. conclusion:the high expression of rrm2 in human cholangiocarcinoma tissue and cell line suggest that rrm2 may be a key gene in initiation and progress of cholangiocarcinoma.

Abstract:objective:to observe the effects of recombinant resistin (rr) on proliferation, invasion, metastasis and matrix metalloprotease-2(mmp-2) expression of human cholangiocarcinoma cell line qbc939. methods:mtt and transwell were used to analyze the effects of different concentrations of rr on proliferation,invasion and migration of qbc939 cells;the expression level of mmp-2 protein in qbc939 cells was examined by western blot. results:rr significantly promoted the proliferation of qbc939 cells when the concentration of rr was higher than 10 ng/ml(p < 0.01),the effect was dose-dependent to some extent. after treated with 10, 50, 100 ng/ml rr for 24 h,the capacity of invasion and migration of qbc939 cells were significantly stronger in rr group than in control group(p < 0.01). the expression of mmp-2 protein was up-regulated significantly in qbc939 cells treated with resistin for 24h. conclusion:rr significantly increases the invasion and metastasis of qbc939 cells in vitro, and the possible mechanism may be related with up-regulation of mmp-2 level.

Abstract:objective:to investigate the effect of silencing cd147 expression on proliferation and invasion of human laryngeal carcinoma cell line hep2. methods:previously prepared recombinant plasmids psilencer-shrna-control, psilencer-shrna1 and psilencer-shrna2 were transfected into the hep2 cell. the rna interference efficiency of cd147 was determined by real-time pcr and western blot. the proliferation was measured by mtt assay. the transwell test was used to detect the invasion ability of the hep2 cells, and psilencer-shrna-control transfected hep2 and normal hep2 were used as control groups. results:after the transfection with psilencer-shrna1 and psilencer-shrna2, the mrna and protein levels of cd147 in the hep2 cells were both significantly down-regulated. proliferation was decreased significantly to 67.49%(p < 0.01)and 61.37%(p < 0.01),respectively. their invasive ability were also significantly decreased to 61.80%(p < 0.01)and 73.58%(p < 0.01),respectively. conclusion:the shrna down-regulated the expression of cd147, inhibited the proliferation and invasive ability of hep2 cells.

Abstract:Objective:To explore the expression and distribution of heme oxygenase isoforms(HO-1 and HO-2) in liver and spleen of cirrhotic rats. Methods:Liver cirrhosis was induced by the subcutaneous administration of CCL4. Distributions of HO-1 and HO-2 in rat tissues were examined immunohistochemically using polyclonalantibodies. The pathologic changes and expression of HO-1 and HO-2 in rat tissues were observed by light microscope and compared using image analysis system. Results:Compared with the controls, HO-1 positive cells were more abundant and intensive in liver and spleen of cirrhotic rats and mainly located in portal areas of liver and in red pulp of spleen. However, no statistical difference of HO-2 between cirrhotic rats and controls was observed. Conclusion:The HO-CO pathway may be involved in the development of liver cirrhosis and pathogenesis of the portal hypertension.

Abstract:Objective: To elucidate pathophysiology of melanocytorrhagy by investigating morphological alterations of dendrites and intracellular calcium concentration(Ca2+) of human melanocytes induced by hydrogen perioxide(H2O2) in vitro. Methods:Melanocytes were cultured routinely, and 0.1% and 0.05% H2O2 were added to culture medium respectively. Dendrites’ length was observed at 30,60,90,120 and 150 min after treatment,and measured accurately by microscope optical ruler. Intracellular calcium was detected with Flu-3/AM probe by flow cytometry. Results:H2O2 induced a notable retraction of dendrites,of which concentration 0.1% had more potent effect than 0.05%. Intracellular calcium was up-regulated with the inducement of H2O2 detected by Flu-3/AM probe. Calcium peak presented was at 30 min after treatment, which is earlier than the first retraction peak of dendrites at 60 min. Conclusion:The results suggest retraction of dendrites might be related to accumulation of intracellular calcium, which led to failure of reflux of calcium.

Abstract:Objective:To determine whether the effect of TNF-α on caspase8/3 activity in mouse adrenal cortex Y1 cells is mediated by TNF-R1. Methods:The expression of TNF-R1 was down-regulated by short hairpin RNAs(shRNAs) in Y1 cells which expressed TNF-R1 natively. The effect of TNF-α on caspase8/3 activity in native Y1 cells and miRNA-Y1 cells was evaluated by spectrophotometer. Results:Interference efficiency of shRNAs on TNF-R1 expression in miRNA-Y1 cells was 71.25%. The activity of caspase8/3 increased significantly in Y1 cells(P < 0.05) and unchanged in miRNA-Y1 cells after treated with TNF-α. Conclusion:The effect of TNF-α on caspase8/3 activity is mainly mediated by TNF-R1.

Abstract:Objective:To evaluate the function of inflammation index in osteoarthritis by continuous dynamic observation. Methods:Seventy New Zealand white rabbits were randomly and equally divided into an osteoarthritis group and a normal control group. Osteoarthritis was induced by injecting 0.5 ml of 2% papain into left knee joints of rabbit,and the same volume of sterile saline solution was injected into left knee joints control group. The serum IL-1β,TNF-α and IL-6 levels were detected on the 1,2,4,6,8,10 and 12 week after injection. Then the rabbits were sacrificed and the knee joints were harvested and studied in pathohistological as well as Mankin scores. Results:Compared with the control group, degenerated changes were observed in OA group, such as thinner articular cartilage, fibrillation and destroyed cartilage matrix,as well as inflammation, proliferation and degeneration of the synovial tissue. All these symptoms got worse with prolonged observation time. The serum IL-1β,TNF-α and IL-6 levels increased in OA group,which were coincided with Mankin scores. Conclusion:Serum IL-1β,TNF-α and IL-6 levels increased with OA process and indicated the progression of disease.

Abstract:Objective:To investigate if the change of N-terminal Brain natriuretic peptide(NT-ProBNP) level with the effect of cardiac resynchronization therapy(CRT). Methods:A retrospective review on 33 patients with CRT/CRT-D during 2005-2009 was conducted. Patients have been followed up for at least 6 months. The patient whose left end-systolic volume decreased by 15% after implantation was considered as responder to CRT. Results:After the implantation, the level of NT-ProBNP decreased significantly in the respond[(2.90±0.57)ng/L vs (3.38±0.47)ng/L,P=0.001], which was in linear regression relationship with the variation of left ventricular end-diastolic volume(b=-0.499,R2=0.489,P=0.001). However, there was no such linear regression relationship existed in non-responder group. The reduction of NT-ProBNP level more than 13% predicted a responsive CRT(HR=14,P=0.02). Conclusion:The reduction of the plasma NT-ProBNP level more than 13% can predict CRT responsiveness. The change of plasma NT-ProBNP level is valuble to monitor of CRT effect.

Abstract:Objective: To investigate the expression of proproteinconvertase subtilisin/kexin type 9(PCSK9) in the rats myocardium impaired by ischemia-reperfusion. Methods:The left descending coronary artery(LAD) of rats was occluded for 45 min, followed by reperfusion of 6,12, or 24 h, respectively. The levels of creatine kinase(CK), creatine kinase MB(CK-MB) and lactate dehydrogenase(LDH) in serum were measured by automatic biochemical analyzer. The pathological changes in myocardium of rat were examined by HE staining. The expression of PCSK9 in myocardium was detected by RT-PCR and Western blot. Results:The levels of CK, CK-MB and LDH were significantly increased after 6 h and 12 h of reperfusion, and restored to normal level after 24 h of reperfusion. The reperfusion-induced myocardial damage increased in a time-dependent manner. The expression of PCSK9 mRNA and protein were significantly and time-depentently increased by ischemia-reperfusion injury. Conclusion:PCSK9 may be invovled in the myocardial ischemia-reperfusion injury.

Abstract:Objective: To assess left ventricular systolic synchronicity and function with right ventricular out tract(RVOT) pacing or right ventricular apex(RVA) pacing by real-time three-dimensional echocardiography(RT-3DE). Methods:In the acute tests of anesthetized closed-chest beagle canines,acustom designed bipolar screw-in lead was implanted using a steerable delivery catheter guided with X-ray. The bipolar leads were implanted in the right ventricular apex or the right ventricular out tract. The dogs received right ventricular pacing at the rate of(138±33) beats per minute. SIEMENS ACUSON SC2000 with 4Z1c probe was used to perform full volume RT-3DE,16-segmental volume curves were obtained by the on-line LVA software.The end-systolic volume(ESV),end-diastolic volume(EDV),Stroke volume(SV),left ventricular ejection fraction(LVEF),the standard deviation(SD)and maximal difference(Dif) of the time to point with minimal systolic volume(Tmsv)of 16 segments,12 segments(6 mid and 6 basal)and 6 segments(basal)were calculated.Tmsv-SD% and Tmsv-Dif% were used to eliminate the influence of different heart rate. Results:There was no significant difference between sinus rhythm(SR)group and RVOT group(P > 0.05).After RVA pacing,Tmsv-16SD%,Tmsv-12SD%,Tmsv-6SD%,Tmsv-16Dif%,Tmsv-12Dif% and Tmsv-6Dif% were significant worsen than those in SR group(P < 0.01),while LVEF was lower(P < 0.05). Compared with RVOT group,Tmsv-16SD%,Tmsv-12SD%,Tmsv-6SD%,Tmsv-16Dif%,Tmsv-12Dif% and Tmsv-6Dif% were increased,and LVEF was decreased significantly(P < 0.05)in RVA group. Conclusion:RT-3DE is a feasible and reproducible method to quantify left ventricular synchrony and function.RVOT pacing in dogs is associated with more favorable immediate myocardial contraction and mechanical synchrony compared with RVA pacing.

Abstract:Objective:To explore the relationship between carotid intima-media thickness(CIMT) and impaired glucose tolerance(IGT). Methods:One hundred and fifty subjects in IGI group and 130 controls were included. IMT and diameter of bilateral common carotid artery were measured by ultrasound automatically. Some biochemical parameters were measured simultaneously. Results:BMI,FPG,2-hPG,TC,LDL-C,TG and the average CIMT of IGT group were higher than that of control group(P < 0.01). There was no significant difference in sex,age,HDL-C,diameter and CIMT on righ and left sides between two groups(P > 0.05). Conclusion:CIMT increased in people with IGT remarkably, which implies that the onset of atherosderotic lesion in them.

Abstract:Objective:To study the effect of aliskiren, a direct renin inhibitor, on oxidative stress in type 1 diabetic rats. Methods: SD rats were randomly divided into four groups(n=9):control group, aliskiren[30 mg/(kg·day)] treated group,diabetic group, diabetic group treated with aliskiren[30 mg/(kg·day)]. Aliskiren was intragastrically administrated daily for 5 weeks, and the effects were compared among groups. Results:Compared with the control group, the activity of xanthine oxidase was decreased in the diabetic group(P < 0.01), the serum nitric oxide (NO) level was reduced(P < 0.01) and the aortic reactive oxygen species(ROS) were increased in the diabetic group. Compared with the diabetic group, aliskiren treatment improved these parameters(P < 0.05). Aliskiren did not modify the lower weight (P < 0.01), the higher blood glucose or HbA1C levels(P < 0.001) of diabetic group. Conclusion:Our results suggest that aliskiren can increase the antioxidant capability of type 1 diabetic rats and decrease the oxidative stress.

Abstract:Objective:To observe the effect of naloxone(Nal) on cardiomyocyte apoptosis and expression of Fas and FasL following acute ischemia-reperfusion injury in vivo and to explore its mechanism. Methods:Twenty-four SD rats were selected and randomly divided into three groups:sham-operated group(sham group), ischemia-reperfusion group(IR group),IR and treated with naloxone group(Nal group). Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) and Western blotting were used respectively to detect the cardiomyocyte apoptosis and protein expressin of Fas and FasL, and histopathological changes of myocardium were observed by optic microscopy. Results:Numerous apoptotic cardiomyocytes were found and the expression of Fas and FasL were increased significantly in ischemic fields in the IR group as compared to sham group(P < 0.01), apoptosis and the expression of Fas and FasL were decreased significantly in the Nal group as compared with IR group(P < 0.01). Local myocytes lesions were observed in myocardium in the IR group, but the injury was lessened significantly in the Nal group. Conclusion:The expression of Fas and FasL in ischemia-reperfusion myocardium can induce cardiomyocyte apoptosis, and naloxone has the cardioprotective effects in ischemia-reperfusion injury by significantly inhibiting the cardiomyocyte apoptosis induced by Fas and FasL expression.

Abstract:Objective:To explore the effect of asiatic acid(AA) on renal c-Jun N-terminal kinase(JNK) signaling pathway and podocyte in diabetic rats. Methods:Diabetic model was established by injection of streptozotocin. The diabetic rats were divided into two groups: diabetes mellitus group(DM group) and AA treated group (AA group). The normal non-diabetic rats were served as the control group (NC group). After eight weeks, the serum urea nitrogen (BUN), serum creatinine(SCR), the excretion of urinary albumin excretion rate (UAER), the blood glucose, the activity of superoxide dismutase(SOD) and the level of malonaldehyde(MDA) in renal cortex were detected. The structure of podocytes was observed by electron microscope. The expressions of p-JNK and JNK protein in kidney tissue of rats were examined by Western blot. The expression of nephrin protein in podocyte was examined by immunohistochemistry. Results:Compared with NC group, in DM group, the levels of BUN, SCR,UAER and blood glucose were significantly elevated(all P < 0.01); the level of MDA in renal cortex was significantly elevated(P < 0.05); the activity of SOD was significantly reduced(P < 0.05); the foot process width was increased, the glomerulus basement membrane was thickened, the mesenterium mass were increased; the expressions of p-JNK and JNK in kidney tissue were significantly increased(all P < 0.01); the nephrin expression was significantly reduced(P < 0.01). Compared with DM group, in AA group, the levels of BUN,SCR,UAER, and MDA in renal cortex were significantly reduced (all P < 0.05); the activity of SOD was significantly increase(P < 0.05); the abnormal structure of podocyte was obviously improved; the expressions of p-JNK and JNK were significantly reduced (all P < 0.05); the nephrin expression was significantly increased(P < 0.05). Conclusion: Asiatic acid may up-regulate the expression of nephrin in the podocyte of DM rats and inhibit the JNK signaling pathway activation showing the protective effect of diabetic nephropathy.

Abstract:Objective: To observe the adiponectin mRNA expression in subcutaneous and omental adipose tissue of type 2 diabetic patients and their relation with serum adiponectin, body mass index(BMI), waist circumference(WC),waist hip ratio(WHR), blood pressure, fasting plasma glucose(FPG), HbA1c, blood lipids and uric acid(UA). Methods:Twenty-three T2DM patients with obesity, 23 T2DM patients, 33 obesity patients and 23 controls were selected. The adiponectin mRNA expression level in adipose tissue was assayed by RT-PCR. The level of serum adiponectin was measured with ELISA kit. FBG, triglyceride(TG), total cholesterol(TC), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), HbA1c, UA of all subjects were determined and their blood pressure, height, weight, waist circumference, hip circumference were measured to calculate BMI,WHR. Results:①The SBP, DBP, FPG, HbA1c, TG, LDL-C, UA were significantly higher, and serum adiponectin, HDL-C and adiponectin mRNA expression level in both subcutaneous and omental adipose tissue were significantly lower in diabetic obesity group than in obesity group(P < 0.05), the adiponectin mRNA expression level in omental were significantly lower than in subcutaneous adipose tissue in both groups(P < 0.01). ②The FPG, HbA1c, TG, UA were higher, serum adiponectin, HDL-C and adiponectin mRNA levels expressed in both subcutaneous and omental adipose tissue were lower in diabetic group without obesity than in control group without obesity(P < 0.05), the adiponectin mRNA expression level in omental was lower than in subcutaneous adipose tissue in both groups(P < 0.01). ③The BMI,WHR,SBP,DBP,FPG,HbA1C,TG,UA were higher, serum adiponectin and adiponectin mRNA level expressed in both subcutaneous and omental adipose tissue were lower in diabetic obese group than in diabetic group without obesity(P < 0.05). ④The adiponectin mRNA expression level in omental adipose tissue of diabetic obesity group was negatively correlated to BMI,WHR,FPG(P < 0.05), and was positively correlated to serum adiponectin(P < 0.01). Conclusion:T2DM patients with obesity show down-regulated expression of adiponectin mRNA in omental adipose tissue and lower serum adiponectin than control obese group,BMI,WC,WHR,FPG are correlated with adiponectin mRNA expression and insulin resistance.

Abstract:Objective:To observe the effects of Shanggan Granule on the express of Toll-like receptor 3(TLR3) and double-stranded RNA-dependent protein kinase(PKR) mRNA in lung of mice with viral upper respiratory tract infection,and explore its immune regulation mechanism. Methods:ICR mice were randomly divided into normal group,model group,antiviral oral liquid group,ribavirin group,Shanggan Granule high-dose group,Shanggan Granule medium-dose group and Shanggan Granule low-dose group.Mice were infected with influenza A virus (A/PR8/34) venom intranasally and treated by different way,the expressions of TLR3 and PKR mRNA were detected by RT-PCR. Results:Compared with the model group,the levels of TLR3 and PKR mRNA in antiviral oral liquid group,ribavirin group and Shanggan Granule groups were reduced significantly(P < 0.05). Conclusion:Shanggan Granule can reduce the expressions of TLR3 and PKR mRNA in lung of mice with viral upper respiratory tract infection,and this way contribute to the immune regulation of Shanggan Granule.

Abstract:Objective:To evaluate the clinical value of acetic acid and indigo carmine staining in the auxiliary diagnosis of flat gastric lesion. Methods:After conventional electronic endoscopic diagnosis, the patients with flat gastrict lesions were randomly divided into 2 groups,staining group was directly sprayed with acetic acid and indigo carmine, then biopsy, the control group was not stained, determined by the naked eye and conventional biopsy. Results:Two hundred and forty patients were adopted, 120 cases of each group. In the staining group, 18 cases had mucosa discoloration, the detection rate of early gastric cancer and high grade intraepithelial neoplasia was 94.4%(17/18), 79 cases appeared poorly stained, the detection rate of intestinal metaplasia and low grade intraepithelial neoplasia was 91.1%(72/79). In the staining group, 10 early gastric cancers, 25 intraepithelial neoplasia and 58 intestinal metaplasia were found, while in the control group 2 early gastric cancers, 6 intraepithelial neoplasia and 32 intestinal metaplasia were found respectively. Conclusion:Staining with acetic acid and indigo carmine improves the detection rates of early gastric cancer and precancerous lesion in flat gastric lesions, especially in the discoloration and poorly stained areas.

Abstract:Objective:To observe the changes of CD4+CD25+Foxp3+T cells in patients with multidrug-resistant tuberchlosis(TB). Methods:We recruited 31 multidrug-resistant TB(MDR-TB) patients,33 drug-susceptible TB(S-TB) patients and 30 healthy controls. Flow cytometry was used to determined the quantity of CD4+CD25+Foxp3+T cells in the peripheral blood. Results:The mumber of CD4+CD25+Foxp3+T cell in the peripheral blood of patients with S-TB was(2.43±0.69)% of peripheral CD4+ T cell, it was significantly higher than heathy controls [(1.61±0.46)%]and lower than patients with MDR-TB[(3.14±0.59)%]. Conclusion:CD4+CD25+Foxp3+T cells increased in MDR-TB patients significantly, this suggests that CD4+CD25+Fox3+ T cells play an important role in the pathogenesis of MDR-TB.

Abstract:Objective:To explore the positive rate and expression levels of p53 and c-erbB-2 in gastric cancer and their relationship with tumor invasion. Methods:SP immunohistochemistry was used to detect the positive rates and expression of p53 and c-erbB-2 expressed in 139 gastric cancer with different invasive depth of lamina propria(T1a),submucosa(T1b),muscularis propria(T2) and suserosa(T3). Results:The positive rates of p53 in T1a and T1b tumors were lower than T3 tumors significantly(P < 0.05),but the expression levels of p53 hadn′s significant difference among tumors with different invasive depth(P < 0.05). The positive rates and expression levels of c-erbB-2 augmented with increased invasive depth of tumor(P < 0.01). The positive rates of c-erbB-2 were related with p53 (P < 0.05),but the expression levels of c-erbB-2 and p53 hadn′t significant relationship (P > 0.05). Ordinal logistic regression analysis showed that both intensity expression of p53 and low, medium, high degree of c-erbB-2 expression affected the invasive depth of gastric cancer significantly(P=0.001,P=0.010,P=0.000,P=0.000,respectively). Conclusion:The expression of p53 and c-erbB-2 in gastric cancer are not synchronous and p53 mutation contributes to c-erbB-2 expression, but the expression level of c-erbB-2 is not related with p53. The mutation of anti-oncogene p53 maybe related to the origin of gastric cancer,while the invasion of gastric cancer is mostly relate to activation of oncogene c-erbB-2.

Abstract:Objective: To evaluate the treatment effect of radical resection of pulmonary carcinoma by video-thoractomy,video-assisted mini-thoractomy(VAMT) and conventional thoracotomy on non-small cell lung cancer(NSCLC). Methods:The clinical material of 1 035 cases with Ⅰ~Ⅱ stage NSCLC underwent,radical resection by three different surgery ways were reviewed. six hundred thirty -seven patients were taken by conventional thoracotomy, 256 patients were taken by video-assisted mini-thoractomy and 142 patients were taken by video-thoractomy. Results:All 1 035 operations were successful. Pulmonary complication occurred in 93 cases,and cardiovascular complication occurred in 37 cases. No significant post-operative complication was found in other 905 cases. All the cases were undergone grouping standard lymph node cleaning, and the prognosis had no significant difference. The radical resection of pulmonary carcinoma by video-thoractomy had the advantages of trauma, less complication, shorter in-hospital time, rapid immunity recovery. Conclusion:The radical resection of pulmonary carcinoma by video-thoractomy is completely feasible and safe. It has the advantages of less trauma and hemorrhage, rapid recovery, artistic margin and so on. It also has less complication and rapid immunity recovery. Therefore the video-thoractomy has the possibility to obtain a more widespread application in the lung cancer surgery.