Abstract:

Abstract:Objective：To investigate the expression of zinc finger protein 262 （ZNF262） mRNA in normal kidney tissues and kidney tissues of patients with autosomal-dominant polycystic kidney disease （ADPKD） at different stages, and to explore the role of ZNF262 in pathogenesis of ADPKD, Methods： Patients with ADPKD were staged according to glomerular filter rate （GFR）. Imaging observation and routine pathological examination were performed, The expression of ZNF262 mRNA and proliferating cell nuclear antigen （PCNA） mRNA in normal kidney tissues （n=8）, early stage ADPKD kidney tissues （n=4） and advanced stage ADPKD kidney tissues （n=4） was examined by semi-quantitative RT-PCR. The correlation between the expressions of the 2 genes was investigated in all tissue specimens, Results： Expression of ZNF262 mRNA and PCNA mRNA in early and advanced ADPKD kidney tissues was significantly higher than that in normal renal tissues （both P〈0.01）, and that in the advanced stage ADPKD was significantly higher than that in early stage ADPKD （both P〈0.05）. The expression of ZNF262 and PCNA mRNA was highly correlated in the early, advanced ADPKD and normal renal tissues（r1 = 0. 842 6, r2 = 0. 902 1 and r3 =0. 883 5, respectively, all P〈0.05）. Conclusion： The ZNF262 mRNA level is higher in ADPKD kidney tissue than that in normal control and increases with the advancement of ADPKD. The expression of ZNF262 is significantly correlated with the expression of PCNA in the same renal tissues. The expression of ZNF262 mRNA may serve as an indicator in diagnosis of ADPKD and may be used for clinical staging of ADPKD patients

Abstract:Objective：To investigate whether celecoxib （CXB）, a specific COX-2 inhibitor, can inhibit the proliferation of cyst lining epithelial cells through blocking mitogen-activated protein kinase （MAPK） signal transduction pathway. Methods： Primarily cultured cells were treated with different concentrations of CXB （0,2.5 × 10^- 6 , 5 × 10^-6 , 1 × 10^-5 , 2 × 10^-5 , 3 × 10^-5 , 4 × 10^-5 ,5 × 10^-5 mol/L） and the proliferative status was evaluated by BrdU assay. The levels of vascular endothelial growth factor （VEGF） were measured by enzyme-linked immunosorbent assay （ELISA） ; the production of proliferating cell nuclear antigen （PCNA） and phospho-MAPK were measured by real-time reverse transcription-PCR assay;and the expression of PCNA, MAPK and phospho-MAPK protein was detected by Western blotting. Results： BrdU assay revealed that CXB inhibited cell growth in a concentration-dependent manner; the maximum inhibition rate （[63.9± 1.2] %） was found when cells were treated with 2± 10^-5 mol/L CXB for 24 h. VEGF secretion by cyst lining epithelial cell was reduced by CXB in a concentration and time-dependent manner. The mRNA and protein levels of PCNA, phospho-MAPK in CXB-treated group were lower than those in control group （with no CXB treatment）. Conclusion： CXB can obviously inhibit the proliferation of cyst lining epithelial cell and the secretion of VEGF, which might be through interfering with the phosphorylation of MAPK and partly blocking MAPK signal transduction pathway.

Abstract:Objective： To investigate and compare the effect and the possible mechanisms of 2 hypoxia-inducible factors （HIF） 1α and 2α on renal cell carcinoma （RCC） by RNA interference （RNAi） method. Methods： Chemically synthesized HIF- 1α and HIF-2α RNAi were transfected into RCC cell line A498 by liposome transient transfection. Normal proximal tubule cell line HK-2 and A498 were separately treated with medium and liposome as controls. The expressions of VEGF, ET-1, bcl-2 and Ki67 were observed by RT-PCR or Western blot 24 h after RNAi. Cell apoptosis was detected by flow cytometry and fluorescence microscopy. Results： Expressions of HIF-1α and HIF-2α RNAi were higher in A498 than those in HK-2, and the 2 genes were successfully blocked after RNAi. HIF-1α RNAi resulted in a significant decrease of ET-1, bcl-2 and Ki67 expression, with increased apoptosis and necrosis of cells. HIF-2α RNAi only resulted in a decrease of VEGF. Conclusion： Both HIF-1α and HIF-2α play important roles in regulation of angiogenesis; moreover, HIF-1α might also regulate proliferation of tumor cells, possibly through regulating anti-apoptotic genes. It is implied the existance of different transcription regulation functions and gene combining sites between HIF-1α and HIF-2α.

Abstract:Objective：To observe the influences of sodium selenite on expression of p38 mitogen-activated protein kinase （p38MAPK） and peroxisome proliferator-activated receptor γ （PPARγ） in rat mesangial cells line HBZY-1, so as to study the role of p38MAPK and PPARγ in diabetic nephropathy and the mechanism by which sodium selenite prevents diabetic nephropathy. Methods： Rat mesangial cell line HBZY-1 was incubated with high glucose, high insulin, H2O2 and advanced glycosylation end products （AGEs） separately before and after HBZY-1 cells were pre-treated with SB203580 （p38MAPK special inhibitor）or sodium selenite. Cells receiving no stimulation were taken as control. The expression of p38MAPK protein and PPARγ mRNA was detected respectively by immunohistochemistry assay and RT-PCR in all groups and the results were compared. Results： High glucose, high insulin, H2O2 and AGEs all activated p38MAPK, increased phospho-p38MAPK expression and decreased the expression of PPARγ mRNA in rat mesangial cells line HBZY-1. The expressions of phospho-p38MAPK protein was markedly inhibited by sodium selenite, while the expression of PPARγ mRNA was significantly increased by SB203580 or sodium selenite in rat mesangial cells lines HBZY-1 （P〈0.01）. Conclusion： p38MAPK may antagonize the expression of PPARγ in rat mesangial cells lines HBZY-1. Sodium selenite, with a similar effect to the agonist of PPARγ,can obviously increase the expression of PPARγ.

Abstract:Objective：To observe the protective effect of peroxisome proliferator-activated receptor γ （PPARγ） on TNFα- induced injury of the inner medullary collecting duct （IMCD） cells. Methods： Cultured IMCD cells were randomly divided into blank control, TNFα stimulation, PPARγ transfection （pPPARγ-control） and PPARγ transfection ＋ TNFα stimulation groups. Stimulation was induced with 50 ng/ml TNFα for 24 h and mouse wild-type PPARγ plasmid was used to transfect IMCD cells. Cell supernatant MCP-1 or TGFβ1 was detected by ELISA method. Results： IMCD cells transfected with wild-type PPARγ plasmid had high expression of PPARγ mRNA and protein. The contents of MCP-1 and TGFβ1 in the supernatant were similar in blank control group and pPPARγ-control group. Compared with blank control group, TNFα stimulation group had decreased contents of MCP-1 and TGFβ1 in the supernatant （P〈0. 005）, but the contents of TNFα stimulation group were significantly higher than those of PPARγ transfection ＋ TNFα stimulation group （P〈0.05 and P（0. 005, respectively）. Conclusion： IMCD cells over expressing PPARγ have anti-inflammatory and anti-fibrosis effects.

Abstract:Objective：To separate human kidney phosphoproteome by two-dimensional gel electrophoresis. Methods： The phosphorylated proteins from human kidney tissues were enriched with phosphate metal affinity chromatography （PMAC） resin. After being concentrated and desalted, the samples were separated by isoelectric focusing on first dimension and SDS electrophoresis on second dimension. Results： The phosphorylated proteins were successfully extracted from human kidney tissues and were separated by two-dimensional gel electrophoresis. Conclusion.. Phosphoprotein enrichment technique combined with two-dimensional gel electrophoresis is an effective approach to study phosphoproteome, laying a foundation for further investigation of human kidney phosphoproteins

Abstract:Objective：To observe whether bone marrow cells （BMCs） can differentiate into the renal tubular epithelial cells under physiological condition. Methyls： Transgenic C57BL/6 mice labeled with green flouresent protein （GFP） served as bone marrow donors and C57BL/6 mice without fluorescence labeling were divided into 3 groups： normal group （N, n = 15）, total body irradiation group （TBI, n= 15）, and bone marrow transplantation （BMT, n= 15） group. The mice in Group N received no treatment, those in Group TBI were irradiated with lethal dose of γ rays but received no transplant （sacrificed within 1 week after irradiation）, and those in Group BMT were transplanted with BMCs after the irradiation （sacrificed at 58 and 84 days after transplantation）. Blood samples were obtained from all 3 groups for routine analysis and analysis of urea nitrogen and plasma creatinine. Meanwhile, mice kidneys were harvested for H-E staining. The amount of GFP positive BMCs in mice of BMT group was determined by flow cytometry; the distribution and amount of GFP positive cells in renal tissues were observed by immunohistochemistry. Results： There was no difference in the levels of urea nitrogen and plasma creatinine between the 3 groups. Lethal-dose total body irradiation resulted in no significant change in renal histological structure and function, but RBC, WBC and PLT of the mice in TBI group were markedly reduced compared with those in the other 2 groups （P〈0.05）. Green fluorescence was seen in renal tubular epithelium of mice on 56 and 84 days after transplantation by fluorescent microscope and laser scanning confocal microscope. Flow cytometry result showed that GFP positive BMCs were （78. 75 ± 5. 99）% and （79. 58±4.60）% in recipient mice on 56 and 84 days after transplantation, respectively; and immunohistochemistry staining showed that GFP positive cells in renal tubular epithelial of the recipient mice were （4.17±1.66）% and （3. 88±1.35）%, respectively （P〉0.05）. Conclusion： Bone marrow cells may participate in renewal of tubular epithelial cells even without the renal tubular damage.

Abstract:Objective: To investigate the growth inhibition and apoptosis induction effect of vitamin E succinate (VES) on human colon cancer cells and to analyze the modulation of apoptosis-mediator Fas expression in this process. Methods: Human colon cancer cell line LS174T was treated with VES for 12 h, 24 h and 48 h at the concentrations of 5 mg/L, 10 mg/L and 20mg/L. 1-(4,5-dimethylthiazo-2-yl)-3,5-diphenylformazan (MTT) assay was employed to detect the inhibitory effect of VES on the growth of colon cancer cells. Flow cytometry was then used to analyze the cell cycle of the colon cancer cells after being treated with VES and the apoptotic rate was calculated at the same time. To find out whether the Fas protein expression was modulated in this process, Western blotting assay and flow cytometry were used to detect the Fas protein level in whole cell lystates and on cell surface. Results: VES exhibited a significant inhibitory effect on the growth of human colon cancer cells in a doseand time-dependent manner. After being treated with VES at 5 mg/L, 10 mg/L and 20 mg/L for 48 h, the apoptotic rate of LS174T cells rose from 0.90％ to 15.9％, 46.7％ and 64.5％, respectively. Fas neutralizing antibody can significantly block VES-induced apoptosis. After the administration of VES, total Fas protein in whole-cell extracts increased in a dose-dependent manner. The flow cytometry showed that the mean fluorescence intensity rose from 5.43 to 9.88, 13.21 and 18.0 after being treated with VES. Conclusion: VES can induce significant growth inhibition and apoptosis in human colon cancer cells. The modulation of Fas expression is one of the mechanisms involved in this process and may be related to the upregulation of Fas molecule on the cancer cell surface.

Abstract:Objective： To study the interaction between RET （GDNF receptor） with sucl-bingding neurotrophic target （SNT1） and search for the possible downstream substrates or regulatory proteins of RET, so as to reveal the mechanism of downstream signal transduction of RET. Methods： The RET^IC （intracellular domain of RET） was fused to LexA and the product was used as a DNA-binding domain protein. SNT1, SNT1PTB or SNT1APTB （SNT1 without PTB） was separately fused to B42AD and their ptoducts were used as an activation domain protein. The cotransformants were tested by β-galactosidase activity analysis and leucine medium growth analysis was used to study the interaction between SNT1 and RET. Results： Interaction between RET and SNT1 or SNT1PTB was found in yeast, but that between SNT1APTB and RET was not observed. Conclusion： It is confirmed that RET can interact with SNT1 in yeast, and the interaction may be mediated by PTB domain of SNT1.

Abstract:Objective： To prepare pure prokaryotically expressed protein of hyrdC and the monoclonal antibody against hyrdC, and to detect the hyrdC protein in gastric cancer with the prepared monoclonal antibody. Methods： The target DNA sequence of hyrdC was obtained by RT-PCR and subsequently inserted into prokaryotic expression vector pET32a. The TRXhyrdC fusion protein was purified and was used to immunize 6-week-old BALB/c female mice to prepare monoclonal antibodies. The supernants of hybridoma cells were screened by enzyme-linked immunoabsorbent assay（ELISA） and identified by Western blot. The prepared monoclonal antibody was used to detect the expression of hyrdC protein in gastric cancer and adjacent normal tissues by immunohistochemical method （SP）. Results： The sequence of hyrdC ORF was conformed to the reported in GenBank. Two lines of hybrids secreting monoclonal antibodies against hyrdC were established,both belonging to IgG2a subtype. Western blot showed that the prepared monoclonal antibodies specifically combined with human hyrdC protein. Immunohistochemistry （SP） results showed that the expression of hyrdC protein in gastric cancer was significantly higher than that in normal gastric mucous membrane （P〈0. 001）. Conclusion： The monoclonal antibody against hyrdC has been successfully obtained. There are significant differences in the expression of hyrdC between the gastric tumor tissues and normal tissues

Abstract:Objective： To investigate the anti-myeloma cells specific immune activity induced by myeloma protein-pusled dendritic cells（DCs）. Methods： DCs were obtained from peripheral blood（PB） of multiple myeloma（MM） patients after co-culture with GM-CSF,IL-4 and TNF-α. The phenotype of DCs was examined by flow cytometry. Host lymphocytes were stimulated with DCs pulsed with myeloma protein acquired from serum of MM patients on DEAE-celluose; lymphocytes treated with unpulsed-DC served as control. [^3 H]-thymidine incorporation and 4 h ^51 Cr-release assay were used to detect the proliferation and cytotoxicity against different targets（auto-myeloma cells,U266 cells） by T lymphocytes. Results： In vitro proliferation and cytotoxicity of T lymphocytes activated by myeloma protein pulsed-DCs were greater than those of T lymphocytes in the control group（P〈0.01）. T lymphocytes in neither of the 2 groups had notable cytotoxicity against U266 cells. Conclusion： DCs pulsed by myeloma protein can effectively induce specific cytotoxicity of T lymphocytes against myeloma cells

Abstract:Objective： To construct a replicating adenovirus vector CNHK600-p53 carrying p53 gene and investigate its effect on the chemosensitivity of hepatocarcinoma cell line. Methods： Chemotherapy of hepatocarcinoma cells BEL-7404 were carried out using Fluorouracil, Mitomycin, Epirubicin, CNHK600-p53, CNHK600-p53 ＋ Fluorouracil, CNHK600-p53 ＋ Mitomycin, and CNHK600-p53 ＋ Epirubicin, separately. MTT assay was used to evaluate the killing effects after therapy. Resuits： The inhibition rate on BEL-7404 cells was（47±3）% when using 100 μg/ml 5-Fu, was （20±4）% when using 1 μg/ml MMC, and was （73±2）% when using 2.5 μg/ml EPI. The inhibition rates of BEL-740 cells in CNHK600-p53（MOI= 10） ＋ Fluorouracil （100 μg/ml）, CNHK600-p53（MOI=10） ＋ Mitomycin （1 μg/ml）, and CNHK600-p53 （MOI= 10）＋ Epirubicin （2.5 μg/ml） groups were （59±4） %, （44±4） % and （86±2） %, respectively （P〈0.01）. Conclusion： Adenovirus CNHK600- p53 carrying p53 gene can enhance chemosensitivity of BELT404 hepatocarcinoma cell line. Gene therapy using adenovirus CNHK600-p53 in combination with chemotherapy may be a new strategy for hepatocarcinoma treatment.

Abstract:Objective： To develop a method for determination of serum midkine （MK） concentration and to evaluate its value in the diagnosis of hepatocellular carcinoma （HCC）. Methods： Since heparin can bind to MK, a heparin-enzyme linked immunosorbent assay （ELISA） for determination of MK was developed using heparin-antibody sandwich technique. Serum MK levels were measured in 20 healthy adults, 20 patients with benign liver tumors, 20 with liver cirrhosis,and 104 with HCC. Results： The method had a minimum detectable concentration of 15 ng/L and a detectable range of （15-1 000） rig/L, and a linearity was found when the concentration was 〈500 ng/L（r=0.97）. Serum MK levels of the healthy subjects,the patients with benign liver tumor, liver cirrhosis and HCC were （56±11） rig/L,（74±19） ng/L, （89±43） ng/L and （199±87） ng/L, respectively; with significant difference found between HCC group and the former 3 groups （P〈0.05）. The positive rate of MK in HCC group was 83.7% ,significantly higher than those of 3 other groups （P〈0. 001）. Conclusion： The heparin-ELISA method for MK determination has diagnostic value for HCC.

Abstract:Objective：To analyze the consistency between the histopathological results of routine endoscopy and biopsy in observing the gastric mucosa of patients with non-malignant gastroduodenal diseases, and to evaluate the necessity of biopsy following gastric endoscopy. Methods.. From Jan. 2005 to Dec. 2005, 320 patients who received upper gastrointestinal endoscopy and biopsy because of upper abdominal symptoms were included in this study. The patients were selected consecutively according to their disorders diagnosed by macroscopic endoscopy and were divided into 64 groups. The 5 patients in each group had reflux esophagitis, non-atrophic gastritis, atrophic gastritis, gastric ulcer and duodenal ulcer, respectively. Patients in the same group were matched with each other in gender, age and their history of diseases. The results of endoscopy, including the exudation, congestion, erosion, roughness, bile reflux, etc. , were read by 2 experienced endoscopists. The biopsy was performed by an experienced pathologist and pathological variables included active inflammation, chronic inflammation, atrophy, intestinal metaplasia and atypical hyperplasia. The status of Helicobacter pylori （H. pylori） infection was evaluated by rapid urea test, silver staining and histological methods; the result was deemed positive when the results of either 2 tests were positive. Results： Erosion, exudation, roughness, and H. pylori infection were related with active inflammation; erosion and H. pylori infection were related with chronic inflammation; roughness of mucosa was related with atrophy; roughness and H. pylori infection were related with intestinal metaplasia; and obsolete hemorrhage, H. pylori infection, roughness mucosa, and bile reflux were related with atypical hyperplasia. Macroscopic diagnosis rate of atrophic gastritis was 71.9% （46/64） with a false positive rate of 28.2 % （18/64） and a false negative rate of 34.38% （22/64）. Conclusion： Macroscopic diagnosis is indicative to pathological changes of gastric mucosa, but the predictive value is relatively poor, making biopsy and pathological examination necessary in the diagnosis of gastric mucosa disorders during routine endoscopic examination.

Abstract:Objective：To evaluate the effect of percutaneous coronary intervention（PCI） on mitral regurgitation （MR） in patients with acute myocardial infarction （AMI）. Methods, A total of 213 AMI patients were divided into PCI group（n = 87, PCI ＋ medication） and medication group（n= 126,medication） according to the treatments they received. Echocardiographic examination was conducted in patients during admission and 6 months follow-up. Color Doppler was used to determine the degree of MR. Echocardiogram indices included MR degree, left ventricular end-diastolic diameter （LVEDd）,left ventricular end-systolic diameter （LVEDs） ,left ventricular end-diastolic volume （LVEDV） ,left ventricular end-systolic volume（LVESV）, and left ventricular ejection fraction （LVEF）. Results： The overall incidence of MR was 28.6% in 213 patients during admission. The MR incidence in patients with acute inferior myocardial infarction was higher than that in patients with other parts of infarction （34.5 % vs 22.3 %, P〈0.01）. MR was found in 35.4 % patients during an average follow-up of 8.8 months. The MR incidence of PCI group was similar to that during admission（26.4% vs 27.6% ,P〈0.05）, while the incidence in medication group increased significantly than that during admission（43.7 % vs 30.2 %, P〈0.01）. Although the MR incidences were not significantly different between the 2 groups during admission, the incidence of medication group was significantly higher than that of PCI group during 8.8 months follow-up（P〈0.01）. LVEDd, LVEDV and LVESV in PCI group were smaller than those in medication group（P〈0.05） ,while LVEF in PCI group was higher than that in medication group（P〈0.05）. Conclusion： The MR incidence in patients with acute inferior myocardial infarction is higher than that in other patients. MR has an increasing trend as the time of myocardial infarction lasts. PCI can prevent MR through improving blood flow and left ventricular remodeling

Abstract:Objective：To assess the efficacy of total intravenous anesthesia （TIVA） with propofol and remifentanil in cervical vertebra surgery. Methods： One hundred patients receiving elective cervical vertebra surgery were randomly allocated to the following 4 groups：group DR, desflurane 1-2 MAC and remifentanil 0. 15-0.40 μg ·kg^-1 · min^-1 ; group DF,desflurane 1-2 MAC and repetitive boluses of fentanyl 0.05 mg; group PR,propofol 50-100μg · kg^-1 · min^-1 and remifentanil 0.15-0. 40 μg · kg^-1· min^-1 ;and group PF,propofol 50-100μg · kg^-1 · min^-1 and repetitive boluses of fentanyl 0.05 mg. During the anesthesia,the adverse cardiovascular response, the use of vascular active drug, time of extubation and time of directive movement resuming were recorded. The mental state and degree of pain were assessed by a blinded observer using simplified State-Trait-Anxiety Inventory （STAI） and Visual Analogue Scale （VAS） 90 min and 24 h after anesthesia. Moreover, the incidence of nausea and vomiting was also recorded. Results： The adverse hemodynamic response,time of extubation and directive movement resuming in group DR and PR were earlier than those in group DF and PF （P〈 0.05）. The STAI scores of TIVA were significantly lower than those of group DR and DF（P〈0.05） 90 min after anesthesia,but the difference disappeard after 24 h. The incidence Of nausea and vomiting in group DR and DF were significantly higher than those in other groups（P〈 0.05） 90 min and 24 h after anesthesia. There was no difference in VAS scores among all groups. Conelusion： The application of TIVA with propofol and remifentanil in cervical vertebra surgery has stable hemodynamics, quicker and complete awakening, good mental state,and reduced incidence of nausea and vomiting. It is worth recommending in clinical practice

Abstract:Objective：To assess the safety and efficacy of the standard manual compression and 2 arterial puncture closing devices, Angioseal and Perclose, for hemostasis at the femoral artery access site in patients undergoing coronary angiography （CAG） or percutaneous coronary interventions （PCI）. Methods： Totally 366 patients undergoing coronary angiography or PCI were assigned to receive either Angioseal （n= 1287, Perclose （n= 110） or standard manual compression （n= 128）. The efficacy endpoint （immediate hemostasis, successful hemostasis rate, operating time, time of leg immobilization and time to hemostasis） and safety endpoint （vasovagal reflex, major complications, local complications, hematocrit drop, etc. ） were evaluated. Risk factors （gender, age, body mass index, comorbid conditions, antiplatelet agents, and anticoagulant agents, etc. ） were also analyzed. Results： The successful hemostasis rates were similar between Angioseal group and Perclose group, but the rate of immediate hemostasis of Angioseal group was higher than that of Perclose group （P〈0.01）. Time of leg immobilization and time to hemostasis in Angioseal group and Perclose group was shorter than those of manual compression group（P〈0.01）. Both vasovagal reflex （9.3 %） and major complications （3.1 %） were only noticed in manual compression group. The rates of local complications in manual compression group and Angioseal group were both lower than that of Perclose group（P〈0.01）. Post-procedure unfractionated heparin was associated with increased risk of hematoma （OR= 4. 382, P〈0.05）, post-procedure clopidogrel （OR=3. 549, P〈0.01） and interventional procedures （OR=6. 584, P〈0.05） were associated with increased risk of blood oozing, and female gender was associated with a reduced risk of blood oozing （OR=0. 132, P〈0.05）. The successful hemostasis rates were lower in the elderly and those receiving PCI. Post-procedure unfractionated heparin and low molecular weight heparin was associated with longer hemostasis time. Conclusion： Perclose and Angioseal can be an alternative of standard manual compression for hemostasis of the femoral artery access site. Angioseal may be better than Perclose regarding the immediate hemostasis and local complications.

Abstract:Objective：To assess the feasibility and accuracy of real-time three dimensional echocardiography （RT-3DE） in quantitative evaluation of left ventricular （LV） volume and ejection fraction （EF） in patients with ventricular aneurysm and myocardial infarction. Methods： Twenty-three patients with left ventricular aneurysm combined with myocardial infarction were examined by RT-3DE, two dimensional echocardiography Simpson＇s method, and M-mode Teichholz method separately. The following parameters： left ventricular end diastolic volume （LVEDV）, end systolic volume （ESV）, stroke volume （SV） and EF were obtained by each method and the results were compared with those obtained by left ventriculography （LVG）. Results.. The values of LVEDV, LVESV, SV, and EF determined by RT-3DE showed good correlations with those determined by LVG （r= 0.92, 0.90, 0.88, and 0.91, respectively; P〈0. 001）, with the values by RT-3DE slightly smaller than those by LVG（P〉 0.05）. The values of LVEDV, LVESV, SV, and EF determined by Simpson＇s method also showed good correlations with those determined by LVG （r= 0.85, 0.87, 0.86, 0.91, respectively; P〈0. 001） , with the values by Simpson＇s method smaller than those by LVG （P〈0.05 for LVEDV and LVESV）. The values of LVEDV, LVESV, SV, and EF determined by Mmode Teichholz method also showed good correlations with those determined by LVG （r= 0.73, 0. 71, 0.70, 0.65, respectively; P〈0. 05）, with the values by M-mode Teichholz method larger than those by LVG（P〈0.05 or P〈0.01）. Conclusion： RT-3DE is a more accurate for evaluation of LV volume and EF in patients with aneurysm combined with myocardial infarction

Abstract:The increasing prevalence of diabetes worldwide has attracted more and more attention; consequently, diabetes literatures have been on a steady rise over the last decade as sustaining efforts were made to control the disease. This paper, through bibliometric analysis of the 1995-2004 literatures （with a defined amount） from ISI Web of Science and other databases, revealed the amount of literatures, citations, and research focuses of diabetes of different countries, institutions, individuals and journals, reflecting the development in diabetes research over the last 10 years from a bibliometric approach

Abstract:Increasing evidences have shown the existance of a mitochondrial nitric oxide synthase （mtNOS）, which binds to the matrix face of the mitochondrial inner membrane and produces nitric oxide （NO） through a Ca^2＋ sensitive pathway. Under physiological condition, the NO catalyzed by mtNOS regulates mitochondrial oxygen consumption and transmembrane potential via reversible competition with cytochrome C oxidase. The reaction of NO with superoxide anion, which was produced by mitochondrial respiratory chain, yields peroxynitrite. Peroxynitrite irreversibly modifies susceptible targets in mitochondria and induces oxidative and/or nitrative stress. In addition, NO has also been implicated in the programmed cell death. This article reviews the current understanding of mtNOS＇s role in the regulation of mitochondrial functions

Abstract:临床资料患者女,27岁,因孕39＋1周于2005年10月15日入院.患者于18岁时无明显诱因出现构语困难,双手发抖,动作笨拙,吞咽困难,至上海华山医院就诊,经血铜蓝蛋白（CP）测定确诊患有肝豆状核变性（hepatolenticular degeneration,HLD）,给予口服青霉胺,4片,1次/d治疗,其后病情稳定.[第一段]

Abstract:Oxygen is a mandatory for all aerobic organisms. Oxygen-containing free radicals are produced when oxygen is not completely reduced to water in energy-producing oxidation reaction. The radicals may also transform into other reactive compounds through electron transfer and all the compounds with similar functions are referred as reactive oxygen species （ROS）. Increased ROS is known to cause damage to proteins, DNA and lipids. Much evidence showed that changes in partial oxygen pressure, hormone, cytokine and chemical stimulation could increase ROS, and ROS, acting as signaling molecules, mediates cell functions. Hypoxia-inducing factor （HIF）, a key transcriptional factor for most hypoxia-inducible genes, is a heterodimer consisting of 2 subunits. Recent study found that ROS plays an important role in HIF activity regulation under hypoxic and non-hypoxic conditions. This paper reviews the production of ROS and its role in the regulation of HIF activity

Abstract:Objective：To develop and validate an accurate three-dimensional geometrical and mechanical finite element（FE） model of the lumbar L4-L5 segment using a new computer-aided designing （CAD） method, Methods： First, a modified ＂nonseed region segmentation＂ was done to extract the interest region in the CT image and to obtain a binary image, from which the iso-surface of vertebral body was produced by a discretized marching cubes algorithm. Second, ＂best cross-section planes＂ representing the morphologic characteristics of physiological lordosis were used for the initial iso-surface model, forming a ＂nonregular piecewise subspace＇. This subspace and the embedded iso-surface model were subsequently transformed by local affine transforms to a ＂regular subspace＇, in which a surface mesh of high quality was generated quickly. Finally, a reverse transform procedure was employed to restore the original three-dimensional （3D） image of the lumbar surface mesh of lumbar L4-L5. All coordinate dada of nodal points and message of triangular patches of the surface model were then subjected to ANSYS for the three-dimensional FE mesh construction. An accurate 3D non-linear FE model of lumbar motion segment （L4-L5） was developed and validated against published data. Results： The constructed FE model of lumbar L4-L5 consisted of 94 794 solid elements, 1196 link elements, 1170 shell elements, 768 target elements and 464 contact elements, and included geometrical, material and contact non-linearities. The predicted results of lumbar L4-L5 segment were closely correlated with published results of experimental biomechanics in terms of axial displacement, segment rotation and intradiscal pressure under similar load condition. Conelusion： Based on advanced algorithm,this constructed surface model of L4-L5 segment is capable to perform whole digitalized binary image extraction and reconstruction of the lumbar surface with excellent simulation results

Abstract:Objective：To develop a compound staining method showing collagenic, elastic fibers and melanocyte in skin tissues. Methods： A staining method combining Masson Fontana silver nitrate, Ponceau S, Picric acid and Victoria blue （MFSNP-PA-VB） was used in the present animal skin experimental. Results： The melanin granule was black in the skin melanocyte, collagenic fibers were red, the red blood cells at hemorrhagic focus were yellow, and the elastic fibers were green by this MFSNP-PA-VB staining method. Conclusion： This compound staining method has multicolor and fine contrast, and can better show melanins, the red blood cells at hemorrhagic focus, and collagenic and elastic fibers

Abstract:Objective：To establish a real-time fluorescent quantitative PCR method for determining the BK virus（BKV） level in renal transplant recipients,and to evaluate its clinical application. Methods： Plasmids containing part of BKV VP1 gene conservative region were constructed as external standards, and a TaqMan probe technique was used to establish a quantitative method for determination of BKV. Urine and peripheral blood（PB） samples from 112 renal transplant recipients were assayed for BK virus levels, and the results were compared with those of 40 healthy controls. Results： The real-time fluorescent quantitative PCR assay established in this study had good sensitivity, specificity, and reproducibility. The minimal detectable level was 8 × 10^2 copy/ml, intro-experiment variation was 0. 54 %-3.78 %, and intra-experiment variation was 0.62 %-4. 58 %. BKV was detected in 27.7% urine samples and 11.6% PB samples from patients. The median level of BKV in urine and PB were 8.2 × 10^4 copy/ml and 2.4 × 10^3 copy/ml, respectively. BKV positive rate of 40 healthy population in urine and PB samples were 2.5 % and 0 %, respectively. The positive rate and level of BKV in renal transplant recipients were both significantly higher than those in normal cohort （both P〈0.01）. The positive rate of BKV in urine samples were significantly higher than that in PB samples（P=0.02）, but the BKV load in urine samples was not related to that in PB samples. Conclusion： The real-time fluorescent quantitative PCR assay in this study is simple, reliable, and precise, which lays a foundation for future study of the relationship between BKV infection and renal graft loss

Abstract:Objective： To establish an acute lung injury model by seawater aspiration in rats. Methods： Twenty-four rats were randomly allocated into 2 groups： control group （n= 9） and model group （n= 15）. The acute lung injury model was established in rats by aspirating 2 ml seawater/kg body weight after 9-10 seconds of breath occlusion. The aspiration was repeated by the same means 2 rain later. Rats in control group aspirated no seawater. Arterial blood gases and respiratory rates were observed before aspiration and 10 min, 30 rain, 1 h, 2 h and 4 h after aspiration. The survival times of animal were also recorded. The rats were sacrificed 4 h after aspiration; the lung index, total protein （TP） contents, white blood cell （WBC） count in bronchoalveolar lavage fluid （BALF）, and the pulmonary histopathology were observed. Results： Compared with control group, model group had a significantly increased respiratory rate and a decreased PaO2 （P〈0.05）,which subsequently rose again and remained at a lower level. The model group also had a markedly increased lung index, TP contents and WBC counts （P〈 0.05）. Obvious edema, alveolar atrophy and inflammatory infiltration were found in model rats. Conclusion： An acute lung injury model by seawater aspiration has been successfuly established

Abstract:目的:构建人B型钠尿肽(BNP)表达质粒,用pET原核表达系统制备重组抗原TRX-BNP融合蛋白.方法:根据GenBank中检索到的人BNP成熟蛋白序列编码,设计合成编码BNP-32蛋白的DNA,并分别在5'端和3'端设计EcoR I和HindⅢ酶切位点,经聚合酶链反应(PCR)、构建重组质粒pET32a.BNP,并在BL21(DE3)经IPTG诱导表达,Ni-NTA亲和层析制备TRX-BNP.通过DNA测序、融合蛋白的相对分子质量分析及Western印迹来鉴定TRX-BNP质粒及表达的BNP抗原的正确性.结果:DNA测序结果表明,所获得的BNP基因编码序列符合序列设计的要求,正确插入设计位点,重组质粒pET32a.BNP构建成功.IPTG诱导后SDS-PAGE电泳显示有特异性蛋白TRX-BNP表达,蛋白相对分子质量21 400,Ni-NTA亲和纯化,在UVP凝胶成像分析系统上作定量分析,BNP蛋白纯度达到95％.Western印迹结果证实TRX-BNP蛋白特异性表达.结论:成功制备TRX-BNP融合蛋白.

Abstract:目的:探讨腹腔镜粘连松解术治疗术后反复发作性粘连性肠梗阻的可行性.方法:33例行腹腔镜粘连松解术患者,均有腹部手术史,术后反复发作性腹胀腹痛、恶心呕吐伴肛门停止排气排便,其中6例有2次手术史,6例曾行开腹粘连松解术.结果:33例患者中30例顺利完成手术,3例中转开腹.腹腔镜手术时间25～90 min,平均(45±10)min,平均出血量约(15±7)ml,1例术中发生肠管损伤,予以腹腔镜下修补.平均术后住院(4.5±0.5)d,术后无并发症.经长期临床随访,31例患者生活质量佳,均未再出现类似术前症状,也无切口疝等其他并发症,2例患者偶有轻微梗阻症状发作.结论:腹腔镜粘连松解术对轻中度粘连引起的肠梗阻具有较好疗效,具有创伤小、腹腔暴露机会少、腹腔干扰轻等优点,能较大限度减少术后腹腔内再粘连.

Abstract:目的:观察通络散结酊(主要组成为天南星、半夏、山慈菇及威灵仙)对大鼠炎症痛模型、骨癌痛模型的镇痛作用.方法:通过Wistar大鼠右踝部注射完全弗氏佐剂(CFA)0.1 ml建立炎症痛模型,造模24 h后应用通络散结酊及扶他林乳剂7d,动态观察大鼠体质量和进食水量、右踝周径及屈/伸踝关节疼痛试验评分的变化.通过Wistar大鼠右胫骨髓腔内注射Walker-256肿瘤细胞3×104建立骨癌痛模型,造模12 d后应用通络散结酊及扶他林乳剂14 d,动态观察大鼠体质量和进食水量、及丙酮刺激缩足反应时间的变化.结果:炎症痛大鼠造模后第2天即出现明显的右踝关节肿胀,屈/伸踝关节疼痛试验评分及右踝周径较空白组明显增加(P＜0.05);与模型组相比,通络散结酊和扶他林乳剂外用均可明显减小大鼠屈/伸关节疼痛试验评分及踝周径(P＜0.05),两种药物作用效果无显著差异.骨癌痛大鼠造模第12天出现缩足反应时间较空白组明显缩短(P＜0.05),通络散结酊和扶他林乳剂外用7 d内均可明显延长骨癌痛大鼠缩足反应时间,较模型组有显著差异(P＜0.05),用药14 d后,通络散结酊仍有很好的效果,而扶他林乳剂效果低于用药7 d时(P＜0.05).结论:通络散结酊对炎症痛及骨癌痛均有镇痛作用,且在扶他林乳剂效果不佳的骨癌痛晚期仍有镇痛作用.

Abstract:目的:探讨AO经皮微创稳定系统LISS钢板治疗胫骨近端复杂骨折的方法和临床效果.方法:21例胫骨近端复杂骨折的患者采用AO经皮微创稳定系统LISS钢板治疗.其中男16例,女5例,年龄45～72岁.致伤原因为交通事故伤17例,建筑施工伤4例.其中7例为Ⅱ度开放伤,14例闭合伤.3例多发伤,6例多发骨折.¨例胫骨多节段骨折,骨折分型按AO分型C型17例,SchatzkerⅥ型4例.结果:本组21例伤口全部一期愈合,全部病例随访3～19个月,平均11个月,手术时间45～160 min,平均80 min,术中无输血,出血100～300 ml.术后患者无发热.未出现小腿筋膜间隙综合征.X线片示骨痂出现时间4周,无短缩以及旋转畸形,仅1例稍向前成角小于5..外观无明显畸形,膝踝关节功能正常.结论:经皮微创稳定系统LISS钢板创伤小,并发症少,骨愈合率高,是治疗胫骨近端复杂骨折的有效方法.

Abstract:目的:合成N,N'-二芳香酰肼类似物.方法:基于HIV-1整合酶结构进行设计,考虑电子效应和空间效应,以不同取代的芳香羧酸等为起始原料,经酰氯化、酯化、肼解等反应方法,进行化学合成.结果:设计、合成了19个二芳香酰肼类衍生物,除LFA3外,其他18个化合物为首次报道.结论:系列新化合物的合成为进一步药物筛选和构效关系研究提供了基础.

Abstract:1 病例资料患者男,47岁,因“反复中上腹胀6个月,右侧胸痛4个月”于2005年2月2日入院.入院体检：右侧胸廓饱满;右侧腋前线第5肋以下压痛,右胸第4肋以下实音;纵隔第2至3肋间向左右两侧增宽,右肺呼吸音减弱,右肺底呼吸音消失,无干湿性啰音.[第一段]

Abstract:1临床资料患儿,女,12岁,因自幼发现心脏杂音,活动后胸闷、气急入院.查体：胸骨左缘第4肋间闻及Ⅳ/6级收缩期喷射样杂音,较粗糙;胸骨左缘2、3肋间闻及Ⅲ/6级收缩期喷射样杂音,P2显著亢进.[第一段]