Abstract:Objective To study the cytotoxic effect of the CdSe/ZnS quantum dots of different diameters on human malignant melanoma cell line A375, A375.s2 and normal human epidermal cell line HaCaT using cell counting kit-8（CCK-8）, so as to study the influence of CdSe/ZnS quantum dots on growth of melanoma cells. Methods Log phase A375,A375.s2 and HaCaT cells were plated into 96-well plates. After a 24 h incubation, the prepared QDs-605 and QDs-545 solutions of different concentrations (162, 81, 54, 40.5, 27, 10.125, 4.05, 2.025, and 1.012 5 nmol/L) were added into the wells. After another 24 h incubation, CCK-8 was added into the well (10 μl/well).Then the cytotoxicities of CdSe/ZnS quantum dots on the proliferation of A375, A375.s2 and HaCaT cells were determined by MTT assay. Results We found that with the increase of QDs-605 and QDs-545 concentrations, the survival rates of A375, A375.s2 cells were decreased. A day after A375, A375.s2 cells were treated with different concentrations of QDs-605 and QDs-545, the cell survival rate was significantly lower than that of the control group(P<0.05,P<0.01); however, different concentrations of QDs-605 and QDs-545 did not inhibit the proliferation of HaCaT cells.Conclusion CdSe/Zns quantum dots have inhibitory effect on the growth of human malignant melanoma cell line A375, A375.s2, and have no noticeable effect on normal human epidermal cell line HaCaT.

Abstract:Objective To verify the interaction between glutamate-ammonia ligase (GLUL) and nuclear localization signal-retinoic acid receptor α (NLS-RARα) protein by yeast two-hybrid and co-immunoprecipitation method. Methods The two plasmids expressing NLS-RARα bait-protein and GLUL protein were co-transformed into yeast AH109 to investigate the interaction in vivo. Tagged fusion protein eukaryotic expression vectors were constructed and co-transfected into HEK 293 cells. Co-immunoprecipitation was used to investigate the interaction between NLS-RARα and GLUL in vitro. Results Positive blue clones were found in the QDO/X-α-gal plate. Eukaryotic expression vectors were co-transfected into HEK 293 cells, then HA-NLS-RARα protein was immunoprecipitated by anti-HA polyclonal antibody, and GLUL-cMyc protein expression was confirmed by Western blotting analysis using anti c-Myc monoclonal antibody. Conclusion The interaction between NLS-RARα and GLUL has been verified by both yeast two-hybrid and co-immunoprecipitation.

Abstract:Objective To investigate the dynamic expression profile of KCNQ1 channel in rat developing pancreas and adult rat islets, so as to provide experimental basis for further analyzing the possible role of KCNQ1 channel in β-cell development and function. Methods The gene expression patterns of embryonic day 12.5 (E12.5), E15.5, E18.5, new-born and adult rat pancreas were compared using the GeneChipRAE 230A. The expressions of KCNQ1 channel subunits in rat pancreas at the above five stages and adult rat islets were also detected and analzyed by RT-PCR. Results The results of genechip showed that the expression of transcription factors related to endocrine cell differention and maturation peaked at E15.5 and E18.5, respectively. The expression of mature β-cell specific genes increased significantly at E18.5, but KCNQ1 and KCNE1 only started to express at E18.5. The results of RT-PCR showed expression of multiple KCNQ1 channel subunits in adult rat pancreas and islet. Conclusion The expression of KCNQ1 channel appears in the later stage of pancreatic development when the endocrine cells undergo further maturation, and has a higher level in adult islets. The expression profile of KCNQ1 channel subunits indicates that KCNQ1 channel might be expressed in mature β-cell and involved in its endocrine function.

Abstract:Objective To screen for human breast cancer sub-cell lines with different metastasis potentials, and to investigate the role of cortactin in proliferation and metastasis of human breast cancer cells. Methods Human breast cancer sub-cell lines with different metastasis potentials were screened by continuous in vitro invasion assays. Ultra-microstructures of sub-cell lines were observed under transmission electron microscope. The proliferation of sub-cell lines was detected by MTT; the cell cycle was observed by flow cytometry; and the migration ability of sub-cell lines was observed by Transwell assay. The mRNA and protein expression of cortactin was examined by immunofluorescence, RT-PCR and Western blotting assay. Results Two sub-cell lines with different potentials of metastasis were obtained through continuous in vitro invasion assay, and they showed essentially identical morphology under the transmission electron microscope. The MTT results showed that proliferation of high metastasis cells was faster than that of low metastasis cells (P<0.05). Flow cytometry demonstrated that the proportion of high metastasis cells in G0/G1 phase was less (\[52.67±3.69\]% vs \[64.46±2.79\]%), and that in S phase was more than that of low metastasis cells (\[30.53±6.19\]% vs \[24.63±2.04\]%). The proliferation index (PI) of high metastasis cells was higher than that of low metastasis cells (\[47.32±3.69\]% vs \[35.53±2.80\]%, P<0.05). Compared with low metastasis cells, the number of high metastasis cells passing through the membrane was significantly more (\[61.46±7.08\] vs \[25.32±4.87\] cells/field, P<0.05). The expression levels of mRNA and protein of cortactin in high metastasis cells was higher than that in low metastasis cells (P<0.05). Conclusion Over-expression of cortactin is closely related to the proliferation and metastasis of human breast cancer cells.

Abstract:Objective To construct the replication-incompetent lentivirus expressing rat TLR4 shRNA, and to observe its inhibitory effect on TLR4 expression in alveolar macrophage and on lipopolysaccharide (LPS)-induced release of IL-1β, IL-6. Methods We designed and constructed four shRNA expressing plasmids with silencing effect, then they were co-transfected into HEK-293T cells with the TLR4 eukaryotic vector constructed previously; the best shRNA was selected to be packed into lentivirus; and the titer was determined. The packed lentvirus was used to infect the alveolar macrophage(NR8383) in presence of LPS, and the LPS-induced IL-1β, IL-6 expression was examined by ELISA. Results The best shRNA was successfully screened out and was correctly inserted into the lentivirus. The titer of the recombinant lentivirus was 2.0×106TU/ml. The LPS-induced expression of IL-1β, IL-6 in the alveolar macrophages was greatly reduced after virus infection(P<0.05). Conclusion We have successfully constructed the recombinant lentivirus expressing rat TLR4 shRNA, which has an satisfactory inhibitory effect against expression of IL-1β, IL-6, paving a way for studying the chronic rejection reaction after rat lung transplantion in vivo.

Abstract:Objective To investigate the effect of Toxoplasma gondii ME49 strain on the apoptosis of mouse placental trophoblastic cells in vitro.Methods Mouse placental trophoblastic cells (concentration of 5×106/ml) were cultured in the different cell culture vessels. The cells were treated for 8 h with different concentrations of Toxoplasma gondii ME49 strain (the concentration of tachyzoites was 2×106 /ml, 4×106 /ml, and 8×106/ml, respectively). FCM was used to examine the apoptosis rates of the placental trophoblastic cells stained with the fluorescent dye of Annexin Ⅴ-FITC/PI; fluorescence microscopy was used to observe the changes of cellular morphology, and Western blotting analysis was used to detect the protein levels of Bax and Bcl-2. Results The trophoblastic cells infected with Toxoplasma gondii ME49 strain showed a higher apoptosis compared to the normal cells(P<0.05), and the apoptosis rates increased with the concentration of tachyzoites in the infected groups. The highest apoptosis rate was 28.37% which was found 8 h after culture with 8×106 /ml tachyzoites. Fluorescence microscope observed that the apoptosis of trophoblastic cells increased with the increase of Toxoplasma gondii. Western blotting analysis showed that the relative expression levels of Bax and Bcl-2 were 1.24±0.05, 1.37±0.03, 1.78±0.04, and 1.15±0.03, 1.09±0.05, 0.97±0.01, respectively, which were significantly different from those of the control group (1.17±0.06, 1.23±0.02, P<0.05). Conclusion Infection with Toxoplasma gondii ME49 strain can promote the apoptosis of mouse placental trophoblastic cells in vitro through up-regulating Bax expression and down-regulating Bcl-2 expression.

Abstract:Objective To express the fusion protein of hOct4 and cell penetrating peptides using prokaryotic expression systems, and to optimize its expression methods and observe the membrane penetrating ability of the fusion proteins. Methods The pET-based prokaryotic expression system was constructed by genetic engineering, and the fusion plasmid was transferred into E.coli BL21(DE3) and Rosetta2（DE3）. The protein was purified by Ni affinity chromatography and identified by Western blotting analysis. The penetrating ability of the Rhodamine-labelled fusion protein was investigated using BJ cells. Results We successfully constructed pET21a(+)-hOct4-11R-His and pET21a(+)-EGFP-11R-His vectors. Fusion proteins hOct4-11-His and EGFP-11R-His were generated by transfering the plasmids into E.coli. The fusion protein was verified by Western blotting analysis and was detected in BJ cells. Conclusion We have successfully generated EGFP-11R-His and hOct4-11R-His fusion proteins, and the proteins can effectively enter the BJ cells and locate around the nuclei.

Abstract:Objective To study the treatment effect of compound ovalbumin proteolysis liquid(COPL) on the hypoproteinemia in dogs and rats. Methods Ligation of common bile duct was performed to induce the cholestasis in 25 female and 25 male beagle dogs. One hundred and fifty male Sprague-Dawley (SD) rats were first given phenobarbital sodium (i.g.) for 1 week, then they were treated with carbon tetrachloride (CCl4, 40%-50%, 1.25 ml/kg, i.p., twice a week) combined with edible alcohol (10%) for 8 weeks. Another 150 male SD rats received exairesis operation of right kidney, and then were treated with adriamycin twice (5 and 3 mg/kg, i.p. on the first and forth week after operation, respectively). The serum levels of serum albumin (SAB), total serum protein (TSP), alanine transaminase(ALT), total bilirubin (TB), creatinine (Cr), and urinary nitrogen (UN) were observed. Animals with SAB level lower than 30 g/L were taken as successful model. The model animals were randomly divided into 5 groups: model control group, 3 COPL treatment groups (low, middle and high dose groups) and non-fat milk group. After treatment for 2 weeks, the animals were killed and the sera were collected for detection of these parameters again. Results The levels of SAB were significantly increased (P<0.05,P<0.01) in meddle and high dose COPL groups and the levels of ALT were significantly decreased (P<0.05,P<0.01) in all the 3 COPL groups in all the 3 hypoproteinemia models. Other indices had no significant changes (P>0.05). Conclusion COPL has satisfactory effect on the hypoproteinemia both in the dogs and rats. Moreover, it has protective effect on the hepatic function.

Abstract:Objective To investigate the therapeutic effect of Tongluo recipe(TLR) on hepatic injury in streptozotocininduced diabetic rats and the possible mechanism.Methods Experimental diabetes was induced by intraperitoneal injection of streptozotocin(60 mg/kg) in male SpragueDawley rats. The study included 5 groups:normal control(CN) group,diabetes mellitus control (DM) group, lowdose TLR (TL) group, middledose TLR (TM) group and highdose TLR (TH) group, with 10 rats in each group. Diabetic rats in the latter three groups received 0.5 g/(kg·d), 1.0 g/(kg·d), 2.0 g/(kg·d) TLR, respectively throughout the experiment. The fasting glucose, blood lipid, and serum liver function indices were measured. Twelve weeks after treatment, rats were sacrificed and the liver tissues were collected to examine the activities of superoxide dismutase (SOD), catalase(CAT), glutathione peroxidase (GSHPx), and malondialdehyde (MDA). The ultrastructure of liver tissues was observed through transmission electron microscope. Results The serum AST, ALT and AKP levels of diabetic rats were increased 12 weeks after intraperitoneal injection of streptozotocin (P<0.01), and electron microscope observation showed characteristic pathological changes of diabetes. TLR treatment (middle and high dose) obviously decreased serum AST,ALT levels(P<0.05)and ameliorated ultrastructure changes of liver tissues: glycogen granules decreased in cytoplasm, mitochondrial cristae became distinct and electron density decreased, appearance of smooth and rough endoplasmic reticulum became nearly normal, and the interstitial collagenofiber decreased. The SOD and CAT activities were significantly decreased and MDA levels were increased in liver tissues of diabetic rats (P<0.01). Twelve weeks of treatment with TLR obviously improved SOD activity (at all three doses, P<0.01) and decreased the levels of MDA (middle and high dose, P<0.01). Conclusion Our data suggest that TLR can improve hepatic injury in streptozotocininduced diabetic rats, probably through increasing activities of antioxidant and depleting lipid peroxidation production.

Abstract:Objective To study the inhibitory effect of nitric oxide-donating aspirin（NO-ASA） on prostatic cancer growth and angiogenesis in subcutaneously-injected tumor in mice. Methods The tumor-bearing mice were subcutaneously injected with NO-ASA, and the tumor growth was observed. ELISA was used to assay the expression of VEGF in mouse sera; real-time PCR was used to examine the expression of VEGF mRNA in tumor tissues; immunohistochemistry was used to detect CD34 expression and to quantify the number of microvessels in the tumor; and Hen’s egg test chorioallantoic membrane (HET-CAM) was employed to detect the in vitro angiogenesis. Results NO-ASA inhibited the prostate cancer growth in mice. The expression of VEGF was greatly decreased in the serum and tumor tissues, and CD34 expression was significantly decreased in the tumor tissues compared with the control group (P<0.05, P<0.01). NO-ASA also greatly inhibited the angiogenesis in HET-CAM. Conclusion NO-ASA can inhibit prostate cancer growth and angiogenesis.

Abstract:Objective To investigate the mRNA and protein expression of cyclin E1 (CCNE1) in gastric carcinoma tissues, and to evaluate its relationship with the development and progression of gastric carcinoma. Methods Oncomine (a cancer microarrays database) was used to analyze the expression of CCNE1 in gastric carcinoma. The mRNA expression of CCNE1 was detected in 34 paired carcinoma and adjacent normal tissues by SYBR Green real-time PCR.Meanwhile, tissue chip/tissue microarray (TMA) technique and immunohistochemistry method were adopted to detect the protein expression of CCNE1 in the 34 matched tissues. Results The CCNE1 mRNA expression was significantly higher in the primary carcinoma tissues than that in adjacent normal tissues (P=0.001); the CCNE1 mRNA expression levels in stage Ⅰ+Ⅱand stage Ⅲ+Ⅳ gastric cancer tissues were both significantly up-regulated (P=0.042,P=0.016). The positive rate of CCNE1 protein was significantly higher in primary carcinoma tissues (41.2%, 14/34) than those in the tumor-adjacent normal tissues (0, 0/34) and gastritis tissues (0, 0/34, P=0.01). CCNE1 protein expression was significantly different in gastric cancer tissues of different differentiation degrees, invasion depths, and lymph node metastasis statues (P<0.05). Conclusion Up-regulated expression of CCNE1 may contribute to the pathogenesis and progression of gastric carcinoma,and detection of CCNE1 may provide reference for diagnosis and prognosis of gastric carcinoma.

Abstract:Objective To study the expression of human giant larvae-1 (Hugl-1) in ovarian carcinoma and its clinical significance. Methods Hugl-1 mRNA expression in 31 ovarian cancer and the corresponding adjacent tissues was examined by real-time fluorescence quantitative RT-PCR and in situ hybridization. Moreover, analysis was done while taking into consideration of the clinicopathologic parameters of ovarian cancer. Results The expression of Hugl-1 mRNA in ovarian cancer tissue was significantly higher than that in the corresponding adjacent tissues (\[0.051±0.029\] vs \[0.026±0.043\],P<0.01); and the expression was not related to patient age, serum CA125 status, pathological characteristics or clinical stages of tumors. Conclusion Hugl-1 may play a role in the development and progresssion of ovarian cancer.

Abstract:Objective To reconstruct a three-dimensional model of the human thoracic aorta, so as to numerically simulate and study the pulsating blood flow in human thoracic aorta. Methods A three-dimensional reconstruction of the human thoracic aorta arch was obtained using CT scan imaging on a human aorta by Mimics, a software for image processing. The numerical simulations were obtained based on the principle of computational fluid mechanics and hemodynamics. Results The distribution of velocity, pressure, and path of the blood flow in the aortic arch of normal people were calculated at a given pulsating and parabolic initial cycle. Our numerical results demonstrated that the blood velocity of distal end was greatly faster than that of proximal end on the interface between branch and aortic arch. There was an obvious pressure gradient between the inner wall and outer wall of blood vessels. The areas where the blood velocity and pressure changed greatly were consistent with the location of the thoracic aortic dissection, especially the area 2-3 cm below the left subclavian artery, suggesting that the blood pressure and velocity may greatly influence the progress of aortic dissection. Conclusion Numerical simulation and modeling based on CT scan data can help to study bio-fluid mechanics. At the same time, the numerical simulation of blood flow will greatly help the diagnosis and treatment of the aortic dissection and arteriosclerosis.

Abstract:Objective To study the antifungal activities of triterpene glycosides derived from the sea cucumber Actinopyga sp. Methods Triterpene glycosides were isolated and purified by silica gel chromatography, reversed-phase silica gel chromatography, Sephadex LH-20, and RP-HPLC from Actinopyga sp. The structures of the triterpene glycosides were elucidated based on spectral data, chemical reactions, and related references. The antifungal activities of the extracts against three strains:Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus were assessed by liquid microdilution method. Results Four triterpene glycosides were identified from Actinopyga sp.; and they included frondoside A (1),pervicoside C (2), holothurin A (3), and holothurin B (4). The MIC80 of compound 2 and compound 3 for antifungal effect were 1-4 mg/L, showing marked antifungal activities.Conclusion Compounds 1 and 2 have been isolated from this halobios for the first time. All the four compounds show considerable antifungal activities, and pervicoside C may be used as a lead compound of antifungal agents.

Abstract:Objective To study the characteristics of peripheral nerve conduction (PNC) in advanced-stage amyotrophic lateral sclerosis (ALS) patients undergoing mechanical ventilation. Methods The sensory conduction velocity(SCV), motor conduction velocity（MCV）, distal motor latency(DML) and amplitude of compound muscle action potential(CMAP) were determined in three ALS patients undergoing mechanical ventilation. The clinical data and PNC characteristics of the patients were analyzed. Results The 12 motor nerves were abnormal in all the three patients, including absence of responsive wave (66.7%,8/12) , decreased amplitude of CMAP(33.3%,4/12), prolongation of DML(16.7%,2/12) and reduction of MCV(16.7%,2/12). The CMAP amplitude of patients gradually decreased with the progression of disease, and finally led to adverse reactions. The SCV and amplitude of sensory nerve action potential were normal in all the three patients. Conclusion ALS patients undergoing mechanical ventilation may have severely damaged motor nerve conduction, and their sensory nerve is generally not affected, with the specific reason remains to be further studied.

Abstract:Objective To explore the rule of voltage change in the reference point of electrocardiograms(ECG). Methods The r and R′ waves on the head-chest lead and routine lead ECGs in 21 patients with right bundle branch block in whole synchronization were compared, and the correlation of voltage change in reference point with the direction of cardiac muscular stimulation of ECGs was analyzed.Results The swing of wave r in head-chest leads was significantly higher than that of routine leads(\[0.16±0.19\] vs \[0.11±0.13\] mV, P<0.01), and the swing of wave R′ in head-chest leads was significantly lower than that of the routine leads( \[0.25±0.21\] vs \[0.35±0.23\] mV, P<0.01). Conclusion Voltage change in reference point of ECG varies with the direction of cardiac muscular stimulation.

Abstract:Objective To use videomimicography to display the skills of retroperitoneal laparoscopic radical nephrectomy in a self control manner, so as to help the beginners to learn the skills quickly. Methods The videos of 147 cases of retroperitoneal laparoscopic radical nephrectomy for renal-cell carcinomas (T1N0M0), which were performed by a surgeon in our department from Jan. 1, 2002 to Dec.31, 2006, were retrospectively analyzed. The first 30 cases were compared with the last 30 cases in terms of operative manipulation, outcomes, problems in operation, and complications after operation. The difficulties in learning retroperitoneal laparoscopic radical nephrectomy from videos were discussed. Results There were no differences between the two patient groups in age, gender, the location or size of the tumors. The bleeding volumes, operation time, frequencies of transferring to open surgery, and addition of Trocar in the first 30 patients were significantly more than those in the last 30 patients (all P<0.01). Results of videomimicography demonstrated that the beginners had a poor anatomic knowledge, the exposure and tissue isolation were not enough, and they had a poor basic skill in manipulating laparoscope. Conclusion Videomimicography can better display the difficulties in retroperitoneal laparoscopic radical nephrectomy in a self control manner, which can help the beginner to understand the operating skills; the method is worth popularizing.

Abstract:TIM (T-cell immunoglobulin and mucin) gene, firstly found on mouse chromosome 11B1.1, is expressed by various kinds of immune cells, especially antigen presenting cells. TIM is involved in both innate immunity and adaptive immunity. Some TIM proteins can bind different ligands, exerting dual influence on T cells. The abnormal expression of TIM is associated with many kinds of diseases, making TIM a new target for pharmacotherapy. The relation of TIM and Tregs has cast new lights on transplantation immune-tolerance. This paper focuses on the immune regulation function of TIM family.

Abstract:1 临床资料 患者，女，27岁。因孕21+6周，右下腹痛伴不规律腹胀4 d就诊。入院前4 d出现右下腹痛，逐渐加重，伴下腹部不规律阵发性胀痛，无阴道流血、流液。2 d前于外院行B超示子宫右侧一约17 cm大小等回声团块，腹水最深4 cm；CA125、AFP轻度升高。诊断为“晚期先兆流产，盆腔肿瘤”，给予抑制宫缩治疗，症状未改善。急诊入我院后查体：右下腹压痛，可及一直径约15 cm包块，腹部移动性浊音阳性。产检：胎心162次/min，有宫缩，质弱，胎膜未破。B超示：子宫右后壁见15.5 cm×10.8 cm×11.6 cm囊实性团块，子宫直肠凹少量积液。CA125 104 U/ml，CRP 32.9 mg/L，AFP、CEA、CA199、CA15-3、SCC均未见异常。急查血常规：Hb 81 g/L。入院诊断：（1）孕21+6周，先兆早产；（2）盆腔包块待查：右侧卵巢肿瘤？给予保胎、对症治疗。入院后第3天晨，患者诉腹痛加重，复查血常规提示Hb降至64 g/L，B超提示盆腹腔积液增多，考虑内出血可能，急行剖腹探查术。术中见：腹腔大量积血及血凝块，总量约4 000 ml。右侧卵巢肿瘤包膜有一约5 cm破裂口，内有烂肉样组织流出，有活动性出血。给予输血、补液，行右侧附件切除术。术后病理：肿瘤细胞卵圆形，梭形，核大深染，显著异型，排列成片状旋涡状，部分区域疏松水肿，肿瘤内可见片状出血。免疫病理：VIM（+），SMA（部分+），Ki-67（），ER（+），PR（+），CK8/18（-），EMA（-），S-100（-）。结合形态学初步诊断：（右卵巢）恶性性索间质肿瘤,疑为未分类性索间质肿瘤或纤维肉瘤。术后给予抗炎保胎对症治疗7 d，患者病情平稳，胎儿情况未提示异常。建议患者引产后行化疗，但患者拒绝，要求出院。

Abstract:The green fluorescent protein (GFP) is a bioluminescent protein derived from the Jellyfish Aequorea．As a reporter gene, GFP has been used in a variety of biological researches. A large proportion of the clinical fungal infections are caused by Candida albicans, making it very important for further study. GFP has also been widely used in Candida albicans-related studies. The fusion genes harboring GFP gene can yield expression of the fusion protein, which can emit green fluorescence and can be used for gene expression and protein localization. This paper reviews the application of GFP in the Candida albicans-related studies.

Abstract:Circulating endothelial progenitor cells (EPCs) derived from bone marrow are capable of endocrine, reendothelialization of injured blood vessels and neovascularization of ischemic lesions; besides, they also play a major role in the pathogenesis of atherosclerosis. However, the number and functions of circulating EPCs are greatly declined in patients suffering from cardiovascular diseases. Functions of EPCs are associated with not only pathophysiological factors such as aging, hypertension, diabetes and hyperlipidemia, but also personal lifestyle such as exercise, body weight and smoking, which directly influence the development of cardiovascular diseases. In this review, we mainly focus on the relationship between EPCs and cardiovascular risk factors and the potential roles of EPC therapy for cardiovascular diseases.

Abstract:Objective To establish a colorectal cancer phage-peptide library and to screen for biomarkers for early detection of colorectal cancer. Methods A T7 phage display peptide library was constructed using 30 surgical colorectal cancer specimens from Changhai Hospital, Second Military Medical University. Protein-A/G was used to enrich IgG from control sera as well as colorectal cancer sera. Five biopanning protocols were carried out for enrichment of colorectal cancer-specific phage clones, and 2 000 phage clones were randomly selected. ELISA was used for further screening of clones of different reactivities between the cancer serum and control serum; and the selected clones were subjected to DNA sequencing and the cloned protein function was forecasted by Chilibot for validation. Results (1) The titer of the colorectal cancer phage display peptide library was 3.0×106pfu, with a recombination rate of 60% as showed by PCR identification and a storage capacity of 1.8×106pfu. (2) Of the 18 phage clones selected by ELISA, 12 were cancer-related genes. Conclusion ELISA for screening the recombinant tumor antigen phage display peptide library can be used to discover new differentially expressed antigens; and the selected phage clones expressing antigen might be used for early detection of colorectal cancer.

Abstract:Objective To construct the recombinant lentiviral vector expressing specific shRNA of p120 catenin gene, and to identify the RNA interference efficiency by infecting PANC-1 cells. Methods Three pairs of shRNA targeting p120ctn mRNA were designed, synthesized and ligated into enzyme-digested and linearized pGCSIL-GFP lentiviral vectors. The recombinant lentiviral vectors were co-transfected with the pGC-FU-p120ctn-3FLAG containing p120ctn gene into 293T cells after identification by PCR and sequencing analysis. Western blotting analysis was applied to select the most effective shRNA. Recombined lentivirus was packaged and the concentration of the virus titer was measured. Real-time PCR and Western blotting analysis were used to identify the interference efficiency after infection of the PANC-1 cells by recombined lentivirus.Results PCR and DNA sequencing analysis confirmed that p120ctn-shRNA-LV was successfully constructed and the concentration of the virus titer was 3×109TU/ml. Real-time PCR showed that expression of p120ctn mRNA was decreased by 82.6% in PANC-1 cells infected with the recombined lentivirus (P<0.05). Western blotting analysis showed that the expression of p120ctn protein was also greatly deceased after infection. Conclusion We have successfully constructed lentiviral vector expressing shRNA of p120 catenin gene, and this lays a foundation for studying the role of p120ctn in invasion and metastasis of pancreatic carcinoma.

Abstract:Objective To optimize the formula and the preparation technique of orally disintegrating tablets of scopolamine hydrobromide(SH).Methods The wet granule compressing method and the direct compressing method were compared to choose the suitable preparation method. The orthogonal design was used to optimize the formula of SH tablets while taking the hardness, disintegrating time and dissolution rate of the tablets as indices, and the optimization was verified. Results The wet granule compressing method was chosen for preparation. The optimized formula was composed of mannitol 40%,MCC 35%,lactose 0,and CMS-Na 10%. And the hardness, disintegrating time and T50 of the optimized tablets were 3.8 kg, 54 s, and 2.47 min, respectively. Conclusion The obtained formula and preparation technique for orally disintegrating tablets of SH is satisfactory; the quality of the prepared tablets can be well controlled．

Abstract:目的 探讨在MCF-7细胞株中雌激素是否影响P2Y2 mRNA的表达。方法 取生长良好的MCF-7细胞,以不同浓度17β-雌二醇(17β-E2)、雌激素受体(ER)拮抗剂ICI 182 780、ERα拮抗剂MPP和ERβ拮抗剂PHTPP作用于细胞,应用实时定量RT-PCR方法测定细胞中P2Y2受体mRNA表达。结果 在MCF-7细胞中,雌激素(1 nmol/L~1 μmol/L)下调P2Y2 mRNA的表达量从对照组的(73.79±8.91)%减少至对照组的(53.68±5.90)%，这种下调作用可以被ER拮抗剂ICI 182 780(1 μmol/L)拮抗,ERα的拮抗剂MPP(50 μmol/L)可以部分阻断雌激素(0.1 μmol/L)的作用,而ERβ拮抗剂PHTPP(50 μmol/L)对雌激素(0.1 μmol/L)的作用无影响。结论 雌激素通过ERα抑制P2Y2 mRNA的表达,这一途径可能参与了雌激素对MCF-7细胞的促增殖作用。

Abstract:目的 观察新诊断2型糖尿病患者血清visfatin水平及罗格列酮、二甲双胍对其的影响。方法 采用ELISA法测定84例新诊断2型糖尿病患者和42例健康对照患者血清visfatin的水平,将糖尿病患者随机分为罗格列酮治疗组和二甲双胍治疗组(n=42),观察罗格列酮、二甲双胍治疗12周后血清visfatin及血糖、胰岛素等临床指标的变化。结果 2型糖尿病患者血清visfatin水平明显高于健康对照组 (P<0.05),与体质量指数（BMI）、空腹血糖(FPG)、糖化血红蛋白（HbAlc）、空腹胰岛素(FIN)、胰岛素抵抗指数（HOMA-IR）呈正相关（相关系数分别为r=0.561、r=0.592、r=0.603、r=0.612、r=0.635,P<0.05）,多元回归分析显示HbAlc、FIN、HOMA-IR是visfatin的独立相关因素（P<0.05）。罗格列酮治疗12周后,患者血清visfatin水平较治疗前显著升高(P<0.05),多元回归分析显示visfatin的变化与其他临床指标变化间无相关性。二甲双胍治疗前后visfatin无明显变化。结论 Visfatin在2型糖尿病的发病中有重要作用,罗格列酮可能增加其在患者血清中的表达。

Abstract:目的 采用正交试验法优选辛夷药材中木兰脂素的微波萃取工艺。方法 以高效液相色谱法(HPLC)测定辛夷药材提取物中木兰脂素的含量作为检测指标,条件是:色谱柱（150 mm×4.6 mm）,流动相为乙腈-水（4060）,检测波长238 nm, 进样量20 μl。采用L9(34)正交试验,考察75%乙醇用量(75~85 ml)、微波萃取时间(600~999 s)和萃取温度(70~80℃) 3个因素。结果 通过正交试验法优选出微波萃取的最佳条件为75%乙醇用量85 ml，萃取时间800 s,萃取温度76℃,该方法提取出的木兰脂素平均含量为6.59%。结论 用正交试验法优选得到的萃取工艺稳定、方法可行,与常规提取法相比,微波萃取技术具有提取时间短、萃取效率高等优点。

Abstract:目的 探讨高压氧(HBO)对舰员重症牙周炎牙龈血流量(GBF)、袋内厌氧菌和谷胱甘肽过氧化物酶(GPx)的作用。方法 90例重症牙周炎患者分为3组，每组30例：HBO组、HBO加洁治组\[HBO+SCA(scaling)组\]和双氧水组(HP组),在治疗前、后即时和治疗结束1个月时用激光多普勒血流仪测定GBF、牙周袋厌氧菌培养、DTNB比色法测定GPx活性,记录牙龈指数(GI)、龈沟出血指数 (SBI)、 菌斑指数(PLI)、 牙周袋深度(PD)、牙周附着丧失(AL)。结果 3组治疗后即时和治疗结束1个月时的GBF、袋内厌氧菌数和GPx与治疗前比较差异均有统计学意义（P<0.01），HBO组和HBO+SCA组治疗后即时和治疗后1个月时的各指标与HP组比较差异有统计学意义（P<0.01）；HBO+SCA组治疗后即时和治疗后1个月时的各指标与HBO组、HP组比较差异有统计学意义（P<0.01），HBO和HBO+SCA组治疗后1个月各指标与治疗后即时比较无差异无统计学意义。结论 HBO对舰员牙周炎有明显治疗作用，HBO加洁治更好，并维持1个月以上，其疗效与抑制致病性厌氧菌，增加GBF和GPx活性有关。

Abstract:查尔酮作为黄酮类化合物家族的一员,具有广泛的生物活性,如抗菌、抗氧化、降血脂、抗炎等作用[1-2]。尤其2’-羟基查尔酮是合成黄酮、黄酮醇以及二氢查尔酮衍生物等的重要中间体,也可用于香料和药物等精细化学品的合成。 目前关于取代查尔酮合成的报道不多,而且反应条件普遍苛刻,反应副产物多,收率低[3]。文献[4]报道了2’-羟基查尔酮的合成方法,我们采用该文献方法以带有不同供电子基的醛和酮缩合,结果重复性不高,且该文报道的产率均为粗产率。为寻找较适合于取代苯甲醛与邻羟基苯乙酮反应的工业化条件,本实验以藜芦醛和邻羟基苯乙酮反应为例（图1）,利用正交试验分析软件最优反应条件,考察在最优反应条件下不同取代的苯甲醛和邻羟基苯乙酮反应的情况。

Abstract:复方大黄颗粒来源于临床经验方,由大黄、黄连、知母、益母草、丹参等七味中药组成,具有补益脾肾、益气养阴、通阳利水，兼以活血之功效,适用于治疗脾肾气虚、淤血、水湿、浊毒内停的糖尿病肾病。为将其研制成为质量稳定、疗效显著、服用及携带方便的现代中药复方制剂,根据方中大黄有效化学成分总蒽醌的理化性质,以蒽醌转移率和出膏率为考核指标,采用综合评分法\[1-3\]对处方中大黄的提取工艺进行正交试验优选。

Abstract:在心血管疾病的众多危险因素中,糖尿病日益受到人们的重视。糖尿病时高血糖、高胰岛素及血脂异常可加重冠状动脉硬化的程度,增高冠心病的发病率\[1\],同时糖尿病亦是非缺血性心力衰竭的重要病因\[2\]。既往大多数有关糖尿病性心力衰竭病理生理变化的基础研究主要集中于血流动力学、神经内分泌、细胞因子和信号转导等方面\[3-4\],对于脂质代谢方面的研究较少。研究已证实糖尿病患者血浆脂质升高的同时,心肌内亦存在脂质蓄积,脂质蓄积可影响心肌对胰岛素的敏感性,使其对葡萄糖的利用能力下降,并可直接损伤心肌细胞,导致心肌细胞出现凋亡\[5\]。新近研究发现在骨骼肌、心肌、脂肪等组织的细胞膜上存在一种参与长链脂肪酸（long chain fatty acid, LCFA）转运的蛋白\[6\],命名为脂肪酸转运体（fatty acid translocase,FAT）,后来发现与人类白细胞分化抗原CD36相同,故简称FAT/CD36。同葡萄糖转运子相似,FAT/CD36主要存在于心肌细胞膜和胞质内\[7\]。关于FAT/CD36在脂质代谢异常时心肌脂质蓄积过程中的作用,目前未见相关报道。本研究观察了2型糖尿病大鼠心肌FAT/CD36的表达量及分布随病程的变化情况,探讨其在心肌脂质蓄积中的作用。

Abstract:儿童肾病综合征是儿童泌尿系统的常见疾病之一，男性比女性发病率高（男女比为3.71）,3~5岁为发病高峰\[1\]，近年来发现肾病综合征发病有年龄趋小之势。本病病因未明，易复发和迁延，年龄越小治疗难度越大。自2006年4月至2009年12月，我们采用低分子肝素联合贝那普利治疗儿童肾病综合征25例，疗效满意，现报告如下。

Abstract:慢性肾病(chronic kidney disease,CKD)目前已成为威胁全球健康的公共卫生问题之一,其患病率呈逐年上升趋势\[1\]。流行病调查提示在美国等发达国家,CKD的患病率约在6.5%~10%\[2\]。多数CKD患者在出现临床症状、肾功能受损或终末期肾病时才被发现,以致错过最佳治疗时机,给家庭、社会带来了巨大的经济负担和精神负担\[3\]。因此开展各种普查及对高危对象的健康宣教,提高公众CKD的知晓率,做到早发现、早诊断、早治疗尤为重要。高血压患者是CKD的高危对象\[4\],约18%的高血压患者最终将出现肾功能下降\[5\]。为此,本研究就社区居民中高血压患者对肾脏及肾脏病相关知识的认知情况进行问卷调查,了解社区CKD卫生服务需求,同时分析其相关影响因素,为进一步开展社区CKD服务提供参考。