Abstract:Objective：To modify the monoantigen DNA vaccine of Mycobacterium tuberculosis with ubiqutin,so as to obtain more potent immune response. Methods: We constructed Mycobacterium tuberculosis MPT64 antigen DNA vaccine (pM) and ubiquitinMPT64 fusion gene DNA vaccine (pUM).The constructed DNA vaccines were intramuscularly inoculated into female BALB/c mice separately. The serum antibodies(including IgG,IgG1,and IgG2a),cytokines (IFNγ,IL4) and cytotoxic T lymphocyte(CTL) response were determined in immunized mice. Results: The IgG level in the pM group was higher than that in the pUM group（P<0.01） and the ratio of IgG2a/IgG1 in the pM group was lower than that in the pUM group（\[2.16±0.3\] vs \[4.48±0.4\],P<0.05）.The IFNγ level was higher (P<0.01） and the IL4 level was lower (P<0.01） in the pUM group than those in the pM group. Furthermore, the pUM group had higher CTL activity than the pM group. It was indicated that the fusion gene DNA vaccine induced weaker humoral immune response but stronger cellular immune response compared to single gene DNA vaccine. Conclusion: The fusion gene DNA vaccine constructed in the present study might be more effective for prevention against tuberculosis than the single gene DNA vaccine.

Abstract:Objective：To clone the Ag85A antigen gene of Mycobacterium tuberculosis and express it in E.coli k802. Methods: The Ag85A gene was amplified from the genome of Mycobacterium tuberculosis by PCR and was inserted into prokaryotic expressing vector pGEX5T to construct pGEX5TAg85A recombinant plasmid. The expression of Ag85A protein in E.coli k802 was induced by IPTG. The expressed protein was purified by affinity chromatography and the antigenicity of the expressed protein was confirmed by Western blotting assay. Results: A band about 0.9 kb in length was obtained by PCR and the recombinant plasmid pGEX5TAg85A was successfully constructed. A new band about 58 000 in length was observed after IPTG induction in E.coli. The relative molecular weight of the expressed protein was consistent with that expected. A single protein band of 58 000 in length was obtained after purification. The expressed protein could be specifically recognized by the sera of patients with tuberculosis patients. Conclusion: The Ag85A gene of Mycobacterium tuberculosis has been successfully cloned and expressed in E.coli k802, which paves a way for further studies on diagnosis and therapy of tuberculosis.

Abstract:1临床资料患者，女，47岁，因“头昏20余年，加重4个月”于2007年7月入院。患者26岁妊娠时出现头昏，在外院就诊，测血压波动于 140～160/90～100 mmHg（1 mmHg＝0.133 kPa），诊断为“妊娠高血压”，产后血压仍高，外院诊断为“原发性高血压”，长期服降压药，近5年服硝苯地平控释片、酒石酸美托洛尔、珍菊降压片控制治疗，血压波动于130～150/90～100 mmHg。2007年3月无明显诱因出现头昏加重，伴恶心、呕吐、四肢乏力，来我院行急诊头颅CT检查，显示左侧侧脑室旁脑梗死，双侧基底节区、半卵圆中心及右侧小脑半球多发性腔隙性脑梗死。患者有时劳累后感胸闷伴心悸，休息后缓解，持续数分钟。7月7日门诊以“高血压原因待查、冠心病”收住入院。查体：血压180/110 mmHg，心率80次/min，律齐；腹部未闻及血管杂音。辅助检查：血、尿常规正常，肝肾功能、血脂正常。心电图：窦性心律，左心室肥大伴劳损。胸片：心脏稍向左扩大，肺、膈未见明显异常。心脏彩超：心脏各房室大小正常，二、三尖瓣少量反流，左室收缩功能正常。肾动脉彩超：右肾动脉狭窄。头颅CT血管成像未见异常。入院后给予硝苯地平控释片30 mg/d；复方利血平片（每片含利血平0.031 25 mg、双肼屈嗪3.125 mg、氢氯噻嗪3.125 mg、氯化钾30 mg）4片/d；酒石酸美托洛尔50 mg/d，福辛普利10 mg/d治疗，血压在170～180/100～110 mmHg。7月12日行冠状动脉＋肾动脉造影检查显示：左冠状动脉左主干、回旋支及右冠状动脉未见明显狭窄，左冠状动脉前降支近中段狭窄10％～30％，第一对角支开口狭窄90％；右肾动脉开口未见狭窄，近中段呈串珠样改变，最大狭窄90％（图1A），左肾动脉未见狭窄。遂对右肾动脉狭窄处行支架置入术，置入Cordis 6 mm×18 mm和7 mm×23 mm支架各一枚，肾动脉狭窄消失（图1B）。术后服硝苯地平控释片30 mg/d，酒石酸美托洛尔50 mg/d，血压降至140/90 mmHg左右。出院诊断：纤维肌性发育不良性肾动脉狭窄，高血压；冠心病，心绞痛。9月25日门诊复查，头昏缓解，劳累后仍感胸闷伴心悸，休息后缓解；血压120/70 mmHg。[第一段]

Abstract:Objective：To investigate the expression of aquaporin 5 (AQP5) and MUC5AC in lung of bronchial asthmatic murine model and the regulatory effect of dexamethasone on the expression.Methods: Fortyeight female C57BL/6 mice were evenly randomized into control group,asthma group and dexamethasone group.The pathological changes of the lungs and the wet/dry weight ratios of the lungs were determined in mice of each group.The expression of AQP5 and MUC5AC mRNA in the lung was determined by realtime polymerase chain reaction (realtime PCR); the AQP5 protein was examined by Western blotting assay; the distribution of AQP5 protein in the pulmonary tissue was studied immunohistochemically; and the content of MUC5AC in the bronchial alveolar lavage fluid (BALF) was determined by ELISA.Results: Compared with the control group,the pulmonary expression of AQP5 in asthma group was obviously decreased (mRNA by\[42.5±3.6\]%,protein by \[64.3±8.2\]%),while the expression of MUC5AC was increased (mRNA by\[93.3±8.1\]%,protein by\[98.3%±7.2\]%) .Dexamethasone significantly decreased the pulmonary expression of AQP5 and MUC5AC compared with the asthma group (P<0.05).Conclusion: The decreased pulmonary expression of AQP5 in asthmatic mice might be related to the elevation of MUC5AC expression in the airway; dexamethasone can decrease AQP5 expression and therefore alleviate the overproduction of airway mucus,pulmonary inflammation and lung edema.

Abstract:Objective：To study the expression of AQP1,AQP3,AQP4 and AQP5 in the pulmonary tissues of mice with acute lung injury (ALI) induced by lipopolysaccharide (LPS) and to clarify the relationship of ALI with the expression of AQP1,AQP3,AQP4 and AQP5. Methods: Forty healthy male C57BL/6 mice were evenly randomized into 5 groups: LPS 4 h group,LPS 6 h group,LPS 8 h group,LPS 10 h group and control group. ALI model was induced with LPS in all the LPS groups. Realtime PCR was used to observe the expression changes of AQP1,AQP3,AQP4 and AQP5 mRNA. Immunohistochemical method and Western blotting assay were used to determine the changes of AQP1,AQP3,AQP4 and AQP5 protein in the pulmonary tissues of all the animals. Meanwhile,measurement of lung wet/dry (W/D) weight ratio and pathological staining were performed in each group. Results: The W/D values of the LPS 4 h,6 h,8 h and 10 h groups (4.39±0.19,4.58±0.17,4.87±0.21 and 5.28±0.16,respectively) were significantly higher than that of the control group (3.99±0.25，all P<0.05). The expression of AQP1 and AQP5 protein in the lung of the LPS 4 h group decreased to (74.1±5.2)% and (70.3±7.1)% that of the control group; the expression of AQP1 and AQP5 protein in the lung of the LPS 8 h group decreased to (45.2±4.4)% and (38.6±8.9)% that of the control group,and the expression of AQP3 and AQP4 had no obvious changes. Conclusion: AQP1 and AQP5 may play important roles in the abnormal ALI fluid transportation and might be associated with the development of pulmonary edema. AQP3 and AQP4 may not participate in the development of pulmonary edema during ALI.

Abstract:Objective：To establish a twodimensional electrophoresis (2DE) gel map of 2 earlystage lung adenocarcinoma tissues with and without RASSF1A expression, so as to screen and identify differential proteins. Methods: Five earlystage lung adenocarcinoma tissues with RASSF1A expression and 5 without RASSF1A expression were screened out by Western blotting assay. The total soluble proteins of the tissue were extracted and were separated by immobilized pH gradient based twodimensional gel electrophoresis to set up the 2DE gel map of the 2 adenocarcinoma tissues. The differentiallyexpressed proteins were analyzed by PDQuest image analysis software and identified by matrixassisted laser desorption/ionization time of flight mass spectrometry (MALDITOFMS); the protein database was searched to further characterized the differential proteins. Results: A wellreproducible 2DE gel map of the 2 adenocarcinoma tissues with and without RASSF1A expression was established and 17 differential protein spots were screened out. Nine of 17 differential protein spots were selected for MALDITOFMS study and satisfactory peptide mass fingerprints were obtained for all the 9 spots. Searching of the protein database revealed 5 candidate proteins and they were: cytochrome b5, 60S acidic ribosomal protein P2, carbonic anhydrase 1, pyrroline5carboxylate reductase 1, and apolipoprotein AⅠ precursor.Conclusion: We have successfully obtained the 2DE gel images of 2 earlystage lung adenocarcinoma tissues with and without RASSF1A expression, and from which we have identified 5 differential proteins,which paves a way for studying the signal transduction pathways involving RASSF1A.

Abstract:Objective：To study the effect of thermochemotherapy on lung cancer and its possible mechanism. Methods: H446 cells were subjected to different thermochemotherapy strategies: 43℃+Paclitaxel (120 μg/L,thermochemotherapy group), 43℃+Paclitaxel (120 μg/L)+SP600125 (20 μmol/L,JNK inhibitor) (thermochemotherapy+SP600125 group), thermotherapy (43℃) group, and Paclitaxel ( 120 μg/L) group; untreated cells served as control. MTT assay was used to measure cell proliferation and Western blotting was used to examine the expression of JNK, pJNK and HSP70 protein. Results: The proliferation rate of cells in the thermochemotherapy group was significantly lower than those in the other 3 groups (all P<0.05). The expression of pJNK was significantly increased in the thermochemotherapy group (P<0.05); SP600125 inhibited the expression of pJNK and the proliferation of cells in the thermochemotherapy+SP600125 group was elevated (P<0.05). The expression of HSP70 in the thermochemotherapy group was lower than that of the thermotherapy group (P<0.05).Conclusion: Thermotherapy can obviously promote the inhibitory effect of Paclitaxel chemotherapy against the growth of lung cancer cell line H446, probably through activating JNK pathway or inhibiting expression of HSP70 protein.

Abstract:Objective：To investigate the expression and distribution of CyPA in nonsmall cell lung cancer (NSCLC) and to evaluate its clinical significance in the development and progression of NSCLC. Methods: The expression of CyPA was examined in 45 NSCLC tissues, 22 paired tumoradjacent tissues and 33 normal pulmonary tissues using tissue microarray immunohistochemistry method. The relationship between CyPA expression in NSCLC and clinic pathological characteristics of NSCLC was analyzed. Results: Expression of CyPA in NSCLC tissues and paired tumoradjacent tissues was significantly higher than that in the normal pulmonary tissues (P<0.05). No significant correlation was found between the expression of CyPA and differentiation and histological types of NSCLC. Conclusion: CyPA is overexpressed in NSCLC tissues, suggesting that CyPA might serve as a biomarker for the development and progression of NSCLC, whose specific role remains to be further studied.

Abstract:Objective：To establish a chronic malignant transformation model of immortalized human bronchial epithelial cell line BEP2D by low dose cigarette smoke condensate (simulating smoking environment).Methods: The chronic dose of cigarette smoke condensate was determined by MTT assay and the colony formation test of BEP2D cells. BEP2D cells were exposed to cigarette smoke condensate once or for multiple times; unexposed cells were taken as control. The malignant tendency of BEP2D cells was identified by antiserum experiment and the malignant features of transformed BEP2D cells were identified by semisolid agar culture. The differentiation ability of BEP2D cells in antiserum experiment and the colony forming rates of BEP2D cells were compared between different groups. Results: The BEP2D cells were exposed to 0.5,1 and 2 μl/ml of cigarette smoke condensate once or for multiple times and were cultured for 25 generations;the differentiation abilities of BEP2D cells(the 25th generation) was significantly different between the cigarette smoke condensate exposed groups (at 0.5,1 and 2 μl/ml) and the normal control group (P<0.05). The cell malignant transformation model was successfully established in the cells of the 38th generation; the cells had multilayer growth and had no contacting inhibition, with chromosome abnormality. The colony forming rates in the semisolid agar culture test was significantly higher in all smoke condensate exposed groups than in control group(P<0.05). The doseresponse relationship showed a good linear correlation (r=0.969, y=42x, P<0.05). Conclusion: The malignant transformation of immortalized human bronchial epithelial cells can be successfully induced by cigarette smoke condensate at 0.52 μl/ml, which offers an ideal model for simulating smoking environment induced chronic malignant transformation.

Abstract:Objective：To establish a nicotinetreatment and withdrawal rat model and to evaluate its characteristics and application through analyzing 3 parameters. Methods：Male SpragueDawley rats, aged 1011 weeks old, were randomly divided into normal saline group(subcutaneous injection of saline \[0.5 ml/kg\] for 6 weeks), nicotinetreated group (injection with nicotine \[0.5 ml/kg, 3 mg/（kg·d）\] for 6 weeks), and nicotinewithdrawn group (injection with nicotine \[0.5 ml/kg, 3 mg/（kg·d）\] for 3 weeks and followed by saline injection for additional 3 weeks). Body weight, food intake, and water intake of animals were recorded during the treatment in 3 groups. The model was evaluated through analyzing body weight, serum parameters and adipose tissue weights.Results：The body weight of rats, as well as the serum levels of triglyceride and insulin, were all decreased after nicotine treatment; the weights of subcutaneous fat, visceral fat and periaortic fat were also decreased. The above indicators increased after withdrawal of nicotine.Conclusion：The established model can be used to study multiple pharmacological effects of nicotine; it can also be used for smoking and smoking cessation related studies.

Abstract:1病例资料患者，34岁，因“停经24+4周，下腹痛10 h”于2006年7月11日收入院，孕2产1，1996年顺产1女，2000年流产1次。月经规律，末次月经2006年1月10日，孕早期过程顺利。10 h前无明显诱因突感下腹疼痛，疼痛进行性加重并向全腹弥漫，伴恶心、呕吐，无发热及阴道流血。查体：体温36.1℃，脉搏105/min，血压 90/60 mmHg(1 mmHg=0.133 kPa)，面色苍白，表情痛苦，心肺听诊无异常，腹肌紧张，肝脾及子宫轮廓触诊不清。WBC 22×109/L，Hb 56 g/L。B超提示：肝脾未见异常，宫内活胎，胎盘位于子宫前壁，胎盘后未见明显液性暗区。腹腔内见大量游离液性暗区，最大液平面达64 mm。立即在B超引导下在最大液平处穿刺，抽出5 ml暗红色不凝血。此时患者突感腹痛如撕裂样加重，随即意识丧失，血压测不出，腹部明显不规则隆起。立即行抗休克治疗并在全麻下行剖腹探查，术中见：腹腔内大量暗红色积血及凝血块，共计约4 500 ml，子宫约孕4个月大小，前壁全层破裂，破口长约10 cm，形态尚规则，胎儿及大部分胎盘组织已流入腹腔，部分胎盘附着于子宫前壁，胎儿已死亡。清理子宫破口周围胎盘、胎膜组织，见胎盘附着处子宫肌壁如鱼肉状，切除部分子宫肌层组织送快速冰冻检查，冰冻提示：(子宫)倾向于胎盘部位滋养细胞肿瘤，良恶性待石蜡切片检查。行子宫修补术，术中、术后共输血4 400 ml。术后病理报告：子宫胎盘部位滋养细胞肿瘤(PSTT)。术后连续复查人绒毛膜促性腺激素（βHCG）均在正常范围，胸片检查提示心、肺、膈未见异常，术后9 d彩超提示：子宫8.5 cm×6.0 cm×8.2 cm，肌层回声不均匀，前壁偏右侧可见大小7.2 cm×4.5 cm×4.0 cm相对高回声团块，边界不清，回声不均，可见多个散在分布的点状、短线状强回声后方伴彗星尾征，宫腔线后移。考虑肿瘤有继续生长侵蚀肌层可能，因患者已无生育要求，术后12 d再次行全子宫切除+双侧卵巢活检术+盆腔淋巴结清扫术。术中未见子宫外转移，标本剖视：子宫孕2个月大小，剖开宫体前壁见5 cm×4 cm×3 cm黑色坏死组织，浸润2/3肌层，宫腔粗糙，病理报告提示双侧卵巢组织未累及。免疫组化：HCG(少量+)，Ki67(少量+)，p53(-)，p16(-)。1周后出院密切随访，血 βHCG均在正常范围，盆腔B超未提示有病灶复发。[第一段]

Abstract:Objective：To investigate the role of HBV subgenotypes B2, C2 in the carcinogenesis, treatment and prognosis of hepatocellular carcinoma (HCC). Methods: HBV genotypes and subgenotypes were detected in 462 HCC patients and 234 chronic hepatitis B (CHB) patients by a multiplex PCR assay, and HCC patients infected with HBV B2 or C2 were followed up for a year after surgical resection, transarterial chemoembolization(TACE) or a combination of both. Results: The HCC patients infected with HBV C2 had a higher chance to receive surgical treatment than those with B2 (P=0.007). Age of 40 years or older (P=0.030), male gender(P=0.000), and viral load (＞10 000 copies/ml) (P=0.017) were the independent risk factors for the carcinogenesis of HCC by using multivariate logistic analysis; however, there was no significant difference in the carcinogenesis of HCC between CHB patients with HBV subgenotypes B2 and C2. Age of 50 years or younger (P=0.044), infection with HBV B2 (P=0.027), and nonsurgical treatment (P=0.000) were the independent risk factors for the recurrence of HCC. Thick trabecular type was more prevalent in HCC patients infected with HBV B2, C2 and genotype mixture (85.7%,71.2% and 75.0%,respectively), and the proportions of histopathological types were not significantly different between HCC patients infected with HBV B2, C2 and genotype mixture. HBV subgenotype C2 was found in all HCC patients with rare histopathological type and subgenotype B2 and mixture were no found.Conclusion: There is no significant difference in the carcinogenesis of HCC between CHB patients with HBV subgenotypes B2 and C2. The HCC patients infected with HBV B2 have a lower chance to receive surgical treatment and are more severe than those with C2. HBV B2 is also closely associated with recurrence of HCC.

Abstract:Objective：To investigate the activation of p38 signaling transduction cascade in renal ischemia reperfusion injury(IRI) and to study the effect of tempol, a free oxygen radical scavenger, on p38 activation. Methods：Male SpragueDawley rats were randomly divided into shamoperation group (n=10), IRI group (n=45) and IRI + tempol group(n=10). Animal IRI model was created by renal pedicle ligation (50 min) of the left kidney along with a contralateral nephrectomy followed by 2 h reperfusion. Rats were sacrificed on 0, 5, 10, 15, 30, 45 min, 1 and 2 h after renal reperfusion. Animals in IRI + tempol group were pretreated with tempol (100 mg/kg) 1 h before undergoing the same protocol as in IRI group; the kidney was harvested after 45 min of reperfusion. Animals in the shamoperation group were subjected to contralateral nephrectomy without renal pedicle ligation and were sacrificed 45 min later. The renal p38 activities of the 3 groups were determined by Western blotting analysis. Malondialdehyde (MDA) content was detected and proinflammatory cytokine TNFα, IL1β levels were analyzed by ELISA. Results： Activation of p38 was observed in the kidney as early as 5 min after reperfusion and reached its peak 45 min after reperfusion and remained to be activated until 2 h after reperfusion（P＜0.05）. The activities of renal p38 in IRI and IRI + tempol group were markedly increased compared with that of the shamoperation group（both P<0.05）. Pretreatment with tempol significantly inhibited IRIinduced p38 activation（P<0.05）; it also decreased MDA activity and TNFα and IL1β levels （both P<0.05）. Conclusion：Our results demonstrate that reactive oxygen speciesmediated p38 activation plays an essential role in IRIinduced renal inflammatory damage in rats, suggesting that inhibition of p38 activation by tempol may be used for prophylaxis and treatment of IRI.

Abstract:Objective：To investigate the effect of hypoxia on the activity of Na+K+ATPase(Na pump) and to understand the distinct functions of high and lowaffinity Na pump during hypoxia of cortical neurons. Methods: Hypoxic condition was mimicked by perfusing cortical slices or culturing cortical neurons with low oxygen solution. Sodium pump current and membrane current of neurons from cortical slices were measured by patchclamp technique in the whole cell mode and the intracellular Ca2+ concentration (\[Ca2+\]i) in cultured cortical neurons was examined by video based motion edge detection system. Changes of Na pump current induced by hypoxia were also examined 4 min after hypoxia in the neurons of cortical slices. Changes of membrane current and \[Ca2+\]i were determined 0, 2, 4, 6, 8 and 10 min after hypoxia with or without Ouabain (Oua, inhibitor of sodium pump). Results: Total sodium pump current were significantly decreased 4 min after hypoxia (\[0.265±0.068\] pA/pF vs \[0.160±0.046\] pA/pF, P<0.01). Membrane current and \[Ca2+\]i were increased in a timedependent manner 10 min after hypoxia (r=0.9803 and r=0.9734, P< 0.01). The effect of hypoxia on membrane current was abolished by tetrodotoxin(TTX,1 μmol/L, a blocker of sodium channel). Oua at 10 μmol/L significantly promoted the hypoxiainduced increase of membrane current and \[Ca2+\]i through inhibiting lowaffinity sodium pump (P<0.05 or 0.01), and Oua at 10 nmol/L significantly depressed the increase of membrane current and \[Ca2+\]i through inhibiting highaffinity sodium pump (P<0.05 or 0.01). Conclusion: Change of sodium pump activity is involved in hypoxic injury in rat cortical neurons. Highaffinity sodium pump is related to the protection of hypoxic injury and lowaffinity sodium pump is related to the hypoxic injury.

Abstract:Objective：To evaluate the in vivo and in vitro effects of Larginine (LArg) on cardiac hypertrophy and the related mechanisms. Methods: (1) In vivo study, rats were divided into shamoperation group, operation group, Larginine (LArg) group, and NnitroLarginine methyl ester (LNAME) group.Coarctation of abdominal aortic artery was performed in rats of the operation group,LArg group and LNAME group. The total protein level in the heart, the heart rate and blood pressure were all determined; cardiac nitric oxide (NO) content was detected by colorimetric method; cardiac ATⅡ content was determined by radioimmunoassay; and the expression of angiotensin converting enzyme (ACE) mRNA was determined by RTPCR. (2) In vitro study, primarily cultured cardiomyocytes were divided into control group, Angiotensin Ⅱ (ATⅡ)treated group, LArg treated group, and LNAME treated group. RTPCR was used to examine the mRNA expression of inducible NO synthase (iNOS), ATⅡ receptor type 1 (ATR1) and ATR2; Western blotting as employed to examine the iNOS protein expression. Results: (1) LArg increased the content of NO in animals of the operation group, decreased the content of ATⅡ, total protein synthesis, ratio of left ventricle mass/body mass, and expression of ACE mRNA; and LNAME could block the above effect of LArg. (2) Compared with ATⅡ group, ATⅡ+LArg higher content of NO, higher expression of iNOS mRNA and protein, and lower ATR1 mRNA level; the above indices were similar between ATⅡ+LArg+LNAME group and ATⅡ group. The expression of ATR2 mRNA was not significantly different between all the groups. (3) Multiple stepwise regression analysis revealed no relationship of regression between total protein level with blood pressure level or the heart rate. Cardiac NO and ATⅡ contents could serve as predictors for total protein level in the heart. (4) Cardiac NO content was negatively correlated with the cardiac ATⅡ content and ACE and ATR1 expression level.Conclusion: (1) The development of cardiac hypertrophy is closely related to cardiac NO and ATⅡ levels. (2) LArg can increase cardiac NO content through enhancing myocardial iNOS expression; NO can reduce ATⅡ production in myocardiocytes and can inhibit the formation of cardiac hypertrophy by reducing the expression of ACE and ATR1. (3) ATR2 does no participate in the above process.

Abstract:Objective：To study the effect of resistin gene on TNFαinduced expression of ICAM1 in ECV304 cells. Methods: Resistin gene was cloned into vector pEGFPC1 to construct pEG/Resi plasmid; the latter was then used to transfect ECV304 cells via liposome.TNFα(20 μg/L) was added to the system 6 h after transfection. RTPCR was used to detect the expression of resistin and ICAM1 mRNA in the cells 18, 30, 42 and 54 h after transfection; meanwhile, the resistin protein levels were determined by Western blotting analysis. ICAM1 protein levels in the supernatants of ECV304 cells were determined by ELISA. Results: Restriction enzyme digestion and sequencing demonstrated that the recombinant plasmid was successfully constructed and was expressed in ECV304. ICAM1 levels in resistintransfected group were consistently higher than that in the nontransfected group in the presence of TNFα. ICAM1 mRNA and protein expression in the transfectedgroup was significantly higher than that in the nontransfected group during the peak periods (30 and 42 h) of resistin expression (P<0.05); ICAM1 mRNA and protein expression increased with the increase of resistin gene expression. Conclusion: Resistin gene can enhance TNFαinduced expression of ICAM1 in ECV304 cells.

Abstract:Objective:To investigate the expression of KAI1 gene in the urothelial cancer tissues and its relationship with the invasion and metastasis of urothelial cancer.Methods: The expression of KAI1 mRNA was detected by realtime fluorescent quantitative(RFQPCR) in urothelial cancer tissues and normal mucosa of urinary tract. The KAI1 protein expression was detected by immunohistochemistry(IHC) method in bladder transitional cell carcinoma tissues and the paired normal mucosal tissues.Results: QRTPCR showed that the average level of KAI1 mRNA in the urothelial cancer tissues was significantly lower than that in the normal bladder tissues (P<0.01); moreover,the increase of pathological grades and clinical stages and the development of lymphatic metastasis were associated with the decrease of KAI1 expression,with significant difference found between the different groups(P<0.05 or P<0.01). The protein expression of KAI1 in the urothelial cancer tissues was significantly lower than that in the normal bladder tissues(P<0.01). The protein expression of KAI1 was decreased with the increase of pathological grades(P<0.05 or P<0.01).We also found that higher expression of KAI1 was associated with superficial invasion (P<0.05) and the presence of lymphatic metastasis (P<0.05).Conclusion: The downregulation of KAI1 gene is associated with differentiation,infiltration,and lymphatic metastasis of urothelial cancer,which might serve as an effective indicator for malignancy,metastasis and prognosis of urothelial cancer.

Abstract:Objective：To discuss the feasibility of distinguishing benign from malignant intraductal papillary mucinous tumors (IPMT) of the pancreas preoperatively based on the clinical and radiological characteristics. Methods: The clinical data of 40 patients with IPMT treated between July 1996 and March 2007 were analyzed. The diagnoses of all patients were confirmed pathologically after surgical resections. Among the 40 patients, there were 17 benign cases and 23 malignant ones. Results: Malignant IPMT patients had a higher frequency of upper abdominal pain and jaundice than benign patients (P<0.05). The sex difference in incidence of IPMT, time of onset, and loss of body mass were not significantly different between the 2 groups. Radiological results demonstrated that the size of tumor and mural nodules in the lesions were larger in the malignant cases than in the benign cases (P<0.05); besides, the malignant cases also had a more dilated main pancreatic duct (MPD)(P<0.05).Mural nodules and thick irregular septum in the cystic lesions were more frequently seen in malignant cases (P<0.05). There was no significant difference in the site preference between the 2 groups. Conclusion: Benign and malignant IPMT have significant differences in their clinical and radiological characteristics, which provides an important evidence for differentiating the biological types of IPMT and helps surgeons on their decision of surgical planning.

Abstract:Objective：To prepare valved mitral stent and to discuss the feasibility of transcatheter implantation of the valved mitral stent in vitro. Methods: We designed a selfexpandable stent made of superelastic NiTi shape memory alloy. The stent was composed of double disks with tubular linkage between them. A valvular ring made of nitinol wire was sutured on the tubular part. Fresh sheep pericardium was crosslinked with a 0.6% glutaraldehyde solution for 36 hours then sutured into a valvular ring; the latter was sutured onto the nitinol selfexpandable stent. Through a sheath across the interatrial septum positioned in the left ventricle of isolated sheep heart via vena cava, the device was delivered into the native mitral valve position. When the left ventricle disk was deployed, it was applied to the mitral annulus by pulling back the external sheath. The waist and the left atrial disk were deployed by pulling the sheath in the annulus and left atrium, respectively. Thus the whole device was deployed in the mitral native. Water was injected into the pulmonary veins and left ventricle to test competence of the prosthetic heart valves．Results: The prepared valved mitral stent could be stably positioned at the native valves. There was no stent migration when it was repeatedly pulled. The two disks sandwiched the native mitral valve with one disk lying in the left ventricle and the other one in the left atrium. The prosthetic heart valves showed satisfactory function without structure damage.Conclusion: This selfexpandable stent is well designed and can be safely implanted in the mitral valve position via transcatheter approach, and the function of the stent is satisfactory.

Abstract:Objective：To prepare valved tricuspid stent and to discuss the techniques of transcatheter tricuspid valve replacement in vitro. Methods: We designed a selfexpandable stent made of superelastic NiTi shape memory alloy. The stent was composed of double disks with tubular linkage between them. A valvular ring made of nitinol wire was sutured on the tubular part. Fresh sheep pericardium was decellularized, treated with 0.6% glutaraldehyde solution for 48 h and then sutured on the valvular ring. Through a 16French sheath positioned in the right ventricle of isolated sheep heart via vena cava, the device was delivered into the tricuspid valve. When the right ventricle disk was deployed, it was applied to the tricuspid annulus by pulling back the external sheath. Then the waist and the right atrial disk were deployed in the annulus and right atrium, respectively, by pulling the sheath. Thus the whole device was deployed in the native tricuspid. Water was injected into the right ventricle to test the competence of the prosthetic heart valves. Results: The prepared valved tricuspid stent could be stably positioned at the native valves. There was no stent migration when it was repeatedly pulled. The two disks sandwiched the native tricuspid valve with one disk lying in the right ventricle and the other one in the right atrium. The prosthetic heart valves showed satisfactory function without structure damage. Conclusion: This selfexpandable stent is well designed and can be safely implanted in the tricuspid valve position via transcatheter approach; and the function of the stent is satisfactory.

Abstract:Objective：To evaluate the validity and reliability of adapted simplified Chinese Version of the Scoliosis Research Society-22 (SRS-22) questionnaire. Methods: The English version of the SRS-22 was translated into simplified Chinese and was subjected to cross-culture adaptation; the final version of the simplified Chinese SRS-22 questionnaire was a consensus reached by an expert committee. Then the simplified Chinese version SRS-22 questionnaire and 36Item Short Form Health Survey (MOS SF-36) questionnaire were used to survey 87 patients with adolescent idiopathic scoliosis who had been surgically treated; 63 patients （72.4％） responded to the SRS-22 questionnaire. The average age of these patients (6 male, 57 female) was (17.7±3.1) years (range, 14.3-23.8 years). Fifty-six of the 63 patients returned the MOS SF-36 questionnaire. The 2 indicators of reliability were Cronbach’s α of internal consistency and intraclass correlation coefficient（ICC）. Concurrent validity of the SRS-22 questionnaire was measured by comparing with domains of the SF-36 questionnaire, which was reflected by Pearson correlation coefficient (r). Results: The Cronbach α values for 4 domains (function/activity，pain，self-image/appearance, and mental health) were all higher than 0.7; and the Cronbach α value for satisfaction of management domain was acceptable. The intraclass correlation coefficients for function/activity，pain，self-image/appearance, mental health, and satisfaction domain were 0.74, 0.78, 0.86, 0.81 and 0.84, respectively, demonstrating a satisfactory reproducibility. For the concurrent validity, 3 domains had excellent correlation, 10 had good correlation, and 21 had moderate correlation. Conclusion: The adapted simplified Chinese Version of the SRS-22 questionnaire has satisfactory reliability and concurrent validity, and might be suitable for post-operation clinical evaluation of Chinese adolescent patients with scoliosis.

Abstract:Receptor tyrosine kinase (RTK),a membrane receptor superfamily with intrinsic protein tyrosine kinase activity,has many members and complicated signal transduction pathways. Activation of RTKs can trigger a series of signal transduction pathways and play essential roles in modulating cell growth,proliferation,differentiation and metabolism through influencing gene transcription and expression. Activation of RTK can also rapidly modulate some cellular functions including the modulation of ion channels. Potassium channels play a critical role in stabilization of membrane potential and regulation of cellular excitability. This review highlights the rapid modulation of potassium channels by RTKs and reviews the recent progress in related research.

Abstract:In recent years, drug resistance of Candida albicans is on a rise in clinical practice. Gene chip technique is the most commonly used means to map gene expression profile in drug resistant Candida albicans. Gene chip can be used to analyze the differentially expressed genes in drug resistant Candida albicans strains induced by various procedures. The drug resistant genes can be identified and their functions can be further studies. The results of gene chip study can also play a role in studying the mechanism of synergism of multiple drug resistance genes.

Abstract:穿膜肽Tat（transcriptional activator protein）是一种带正电荷的短肽,可以协助大分子物质进入哺乳动物细胞。最近的研究表明,多种与Tat融合的物质能够穿过细胞膜而且可以发挥其生物学功能。这项技术对结合物的大小没有严格的限制,即使是原本无法使用的大分子物质也有可能用于治疗疾病。Tat已经在肿瘤、糖尿病、免疫治疗等临床领域展现出很大的研究价值,并且已经取得了不少新的进展,具有广泛的应用前景。

Abstract:在牙颌缺损的修复中，除了牙体色彩外，牙槽和牙龈黏膜的颜色也扮演了重要角色。本文从常用义龈修复材料、牙龈表色系统、牙龈颜色的测量三方面对牙龈色彩相关研究进行回顾和分析。

Abstract:目的：观察替勃龙（tibolone）治疗对绝经后妇女颈动脉和椎动脉内中膜厚度（IMT）和血流阻力指数（RI）的影响。方法:有更年期症状的绝经后妇女138例，随机分为治疗组（替勃龙2.5 mg/d）和对照组。替勃龙治疗前及治疗2年后检测血总胆固醇（TC）、三酰甘油（TG）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C），颈动脉IMT，颈动脉、椎动脉RI。结果:129例完成观察和治疗。替勃龙治疗2年后血TC显著降低（P<0.05），LDL-C和Lp(a)也显著降低（P<0.01），HDL-C虽有降低但无统计学差异（P>0.05）。替勃龙治疗2年颈总动脉IMT下降26.7%（P<0.01）。颈总动脉RI从0.72±0.06下降到0.68±0.06，颈内动脉RI从0.59±0.05降至0.56±0.05,椎动脉RI从0.66±0.06下降到0.62±0.04 (P<0.05)。但颈外动脉RI下降不明显(P>0.05)。结论:替勃龙可降低TC、LDL-C和Lp(a)，颈动脉RI，颈动脉IMT，对动脉粥样硬化有预防作用。

Abstract:准分子激光手术矫正屈光不正，以安全性好、精确度高、损伤小、恢复快等优点，越来越被临床医师所肯定和接受。国内外研究[1-2]表明高度近视患者青光眼的发生率要高于正常人群，因此术后眼压的随访就显得尤为重要。目前广泛使用的非接触式眼压计测量值受角膜厚度等因素的影响，术后眼压的测量值低于真实值[3]。国外不少学者[4-5]为此推算出相应的校正公式以弥补上述缺陷，但效果不很理想。房水流畅系数（C值）与眼压关系密切，它是反映房水在房角流畅程度的指标。目前关于准分子激光手术对房水流畅系数和房角结构影响的研究不是很多。因此，本研究对准分子屈光手术术后猫眼房水流畅系数及房角小梁组织结构进行观察，探讨屈光手术对其的可能影响。

Abstract:目的：探讨颈清扫术后原清扫区域颈淋巴结复发癌的特点及再手术的策略和疗效。方法和结果:回顾分析41例头颈部恶性肿瘤颈清扫术后不同清扫区域颈淋巴结癌复发频率及再手术疗效。各区域发生颈淋巴结复发癌频率Ⅱ区为35.59％（21/59）， Ⅲ区32.03％(19/59)， Ⅳ区15.25％(9/59)，Ⅰ区10.17％(6/59)，Ⅵ区5.08％(3/59)，Ⅴ区1.69％(1/59)，肿瘤直径<3 cm 13例，3~6 cm 19例，>6 cm 9例，首次颈清扫手术后淋巴结癌复发时间1年以内者7例，1~2年者19例，2~3年者7例, 3年以上者8例。颈淋巴结复发癌行单纯肿块切除7例，一个区域8例及两个区域以上者26例再次行颈淋巴结清扫术，其中同时行颈动脉外膜切除术5例、颈动脉切除端端吻合术2例、大隐静脉颈动脉重建术2例。手术并发症发生率15％，围手术期死亡率为0。再次手术后生存6~12个月5例，生存1~2年7例，2~3年10例，3年以上14例，失访5例，3年生存率34.15%，1年以上生存率75.6％。结论:颈淋巴结复发癌发生的部位主要集中在Ⅱ区、Ⅲ区、Ⅳ区和Ⅰ区，尤以Ⅱ区为甚，再次手术很大程度上能延长患者生命。

Abstract:目的：探讨18F-FDG双探头符合线路显像(DHC)对肺部孤立性结节(SPN)的诊断价值。方法:回顾性分析了34例 SPN的18F-FDG DHC及胸部多层螺旋CT检查结果，并结合病理分析。结果:DHC诊断恶性病灶28个，良性病变6个。CT诊断恶性病灶29 个，良性病灶5 个。病理结果提示恶性病灶26个，良性病变8个。DHC诊断SPN的灵敏度 96% ，特异性67%，准确性88%，阳性预测率81%，阴性预测率83%。多层螺旋CT诊断SPN的灵敏度 92%，特异性38%，准确性79%，阳性预测率83%，阴性预测率60%。结论:18FFDG DHC对诊断SPN有临床价值，其特异性、准确性和阴性预测率高于CT。

Abstract:目的：探讨幽门螺杆菌(Hp) 感染及血清同型半胱氨酸水平与老年血管性痴呆及脑梗死的关系。方法:应用ELISA法检测老年(≥60岁)血管性痴呆、动脉硬化性脑梗死及腔隙性脑梗死患者与对照组各40例血清Hp-IgG水平及同型半胱氨酸水平,同时检测血清总胆固醇、三酰甘油、载脂蛋白A、载脂蛋白B水平。结果:老年血管性痴呆、动脉硬化性脑梗死患者血清Hp-IgG、同型半胱氨酸、三酰甘油水平均显著高于腔隙性脑梗死患者及对照组(P<0.01)。各组之间Hp-IgG阳性率无显著差别(P>0.05)。除三酰甘油外，余血脂水平各组之间均无显著差异。结论:血清HpIgG和半胱氨酸水平与老年血管性痴呆及动脉硬化性脑梗死的发病密切相关。

Abstract:鼻咽癌是我国南方地区常见的恶性肿瘤，其发病率近年有增高的趋势。由于其原发部位深而隐蔽，早期症状不典型，临床表现多样化，故误诊误治的报道常见于文献中\[1\]。鼻出血是鼻咽癌的早期症状，而鼻咽纤维血管瘤的主要症状就是鼻出血，如果鼻咽癌外观与鼻咽纤维血管瘤相似，两者又都是以鼻出血为主要症状，那如何有效地鉴别两者，使诊治工作少走弯路，在临床实践中仍是一个值得探讨的问题。下面报告2例鼻咽癌误诊为鼻咽纤维血管瘤的病例，分析其原因并总结改进措施。[第一段]