Abstract:The article presents an overview of the current clinical application of clopidogrel in coronary artery surgery. The viewpoint is that clopidogrel can reduce preoperative and postoperative ischemic events of coronary artery bypass grafting(CABG). With the development of standardized medication and the corresponding preventive technique, it will be of great value to reduce hemorrhage complications and obtain the maximum benefit from clopidogrel’s anti-platelet properties.

Abstract:Cervical arthroplasty after anterior decompression with insertion of a prosthetic total disc replacement has been suggested as an alternate to anterior cervical fusion. It develops quickly during recent years. Currently there are several cervical arthroplasty devices. Each device varies in terms of materials, range of motion and constraint. Early studies suggest that in the short term, the complication rate and efficacy is no worse than fusion surgery. Long-term results have not yet been reported. This review examines the current prostheses as well as discussing issues regarding indications and technique. It is hoped that an improvement of cervical arthroplasty occurs in terms of materials and design as spinal surgeons enter a new times of the management of cervical spine disease.

Abstract:Objective: We have continued previous work in which we demonstrated that #117 and #372 amino acids contrib-uted to the high activities of human CYP2A13 in catalyzing 4-methylnitrosamino-1-(3-pyridyl)-1-butanone(NNK) and aflatoxin B1(AFB1) carcinogenic activation. The present study was designed to identify other potential amino acid residues that contribute to the different catalytic characteristics of two CYP2A enzymes, CYP2A6 and CYP2A13, in nicotine metabolism and provide insights of the substrate and related amino acid residues interactions. Methods:A series of reciprocally substituted mutants of CYP2A6Ile300→Phe, CYP2A6Gly301Ala, CYP2A6Ser369→Gly, CYP2A13Phe300→Ile, CYP2A13Ala301→Gly and CYP2A13Gly369→Ser were generated by site-directed mutagenesis/baculovirus-Sf9 insect cells expression. Comparative kinetic analysis of nicotine 5’hydroxylatin by wild type and mutant CYP2A proteins was performed. Results:All amino acid residue substitutions at 300, 301 and 369 caused significant kinetic property changes in nicotine metabolism. While CYP2A6Ile300→Phe and CYP2A6Gly301→Ala mutations had notable catalytic efficiency increases compared to that for the wild type CYP2A6, CYP2A13Phe300→Ile and CYP2A13Ala301→Gly replacement introduced remarkable catalytic efficiency decreases. In addition, all these catalytic efficiency alterations were caused by Vmax variations rather than Km changes. Substi-tution of #369 residue significantly affected both Km and Vmax values. CYP2A6Ser369→Gly increase the catalytic efficiency via a significant Km decrease versus Vmax enhancement, while the opposite effects were seen with CYP2A13Gly369→Ser. Conclusion:#300, #301 and #369 residues in human CYP2A6/13 play important roles in nicotine 5’-oxidation. Switching #300 or #301 residues did not affect the CYP2A protein affinities toward nicotine, although these amino acids are located in the active center. Ser369 to Gly substitution indirectly affected nicotine binding by creating more space and conformational flexibility for the nearby residues, such as Leu370 which is crucial for many hydroxylations.

Abstract:Objective:To apply reformed AS-PCR, which combined phosphorothioate-modified primers with exo+ polymerase, in single nucle-otide polymorphism discrimination of mitochondrial DNA 10400 locus. Methods: We used the mtDNA 10400 locus to design unmodi-fied and 3′phosphorothioate-modified allele-specific primers for PCR, which was performed using polymerases with and without 3′exonuclease activities. The effects of these primers on primer-extension were evaluated by agarose gel electrophoresis. Results: The unmodified primers were extended by both exo- and exo+ polymerase irrespective of whether the primers were matched or mismatched with the templates. However, the 3′phosphorothioate-modified primers with a terminal mismatch triggered an “off-switch”of exo+ polymerase when compared to exo-polymerase. Conclusion: The“on/off”switch constituted by the combination of 3′phosphorothioate-modified primers with exo+ polymerase is a cost-effective, high-throughput and reliable method for SNP typing, which will be of enormous application in association studies by single nucleotide polymorphism screening.

Abstract:Objective: To observe the effects of atorvastatin on serum matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9(MMP-9), and the tissue inhibitor of metalloproteinase-1(TIMP-1) in the development of chronic heart failure. To investigate the role of atorvastatin in the therapy of chronic heart failure and determine its possible mechanism of action. Methods: Thirty Japanese Big Ear rabbits were randomly selected and divided into 3 groups: sham-operated group(SO group), heart failure control group(HC group) and heart failure atorvastatin therapy group(HA group), with 6, 12 and 12 animals in the respective groups. Volume overloading was produced in the HC group and HA group animals by creating an aortic insufficiency, induced by damaging the aortic valve with a catheter introduced through the carotid artery. After 14 days, abdominal aorta constriction was performed in order to obtain a pressure overload. Six weeks later rabbits in the HA group were administered atorvastatin 3mg. Kg-1.d-1 for 4 weeks, at which time the experiment was terminated. Arterial blood was drawn and serum levels of MMP-2, MMP-9 and TIMP-1 were measured in all groups at the same time using an ELISA method. Results: Structural and functional indicators of chronic heart failure(CHF) were seen in both the HC and HA groups, but atorvastatin significantly reduced the observed effects. The serum concentrations of MMP-2, MMP-9 and TIMP-1 were at low levels in all three groups at the start of the study, with no difference between them(P < 0.05). At the end of 6th week concentrations were significantly increased in the HC and HA groups compared with the SO group(P < 0.05), but there were no differences between the HC group and HA group(P > 0.05). The increased concentrations in HC group continued to the end of the experiment, but values in the HA group were all lower than those in the HC group by the end of the experiment(P < 0.05). Conclusion: Serum concentrations of MMP-2, MMP-9 and TIMP-1 increase significantly during the course of CHF, paralleling the pathological progress of CHF. Atorvastatin benefits CHF, and the decreased serum levels of MMP-2, MMP-9 and TIMP-1 may be one of the drug’s mechanism of action.

Abstract:Objective: To study the morphologic abnormalities of the myenteric plexus in diabetic rats and to explore the mechanism of their effect on gastrointestinal motility. Methods: Forty rats were randomly divided into a diabetic group and a control group, Gastric emptying and small intestine transit rates were measured and histologic and molecular changes in glutamatergic nerves in the ileal myenteric plexus were observed, mGluR5 receptor and EAAC1 transporter changes in the diabetic rats were studied using fluorescence immunohistochemistry and RT-PCR. Results:Eighteen weeks after the establishment of the diabetic rats model, gastric emptying and small intestine transit rates were found to be significantly delayed in the diabetic group when compared with the control group. The density of glutamatergic ganglia and neurons in the ileal myenterie plexus were significantly decreased in the diabetic group when compared with control group(P < 0.05) and the mGluR5 receptors and EAAC1 transporters were downregulated in the diabetic rats(P < 0.05). Conclusion: Decreased glutamatergic enteric ganglia and neurons and decreased mGluR5 receptors and EAAC1 transporters in the intestinal myenteric plexus is one of the mechanisms of diabetic gastroenteropathy in rats.

Abstract:Objective:To find the value of the apparent diffusion coefficient (ADC) of diffusion weighted magnetic resonance imaging of common renal diseases. Methods: There were 30 healthy subjects and 81 patients with renal lesions (56 cases of renal carcinoma, 18 lesions of 12 cases of renal angiomyolipoma, and 21 lesions of 13 cases of renal cysts). Conventional magnetic resonance imaging and diffusion weighted magnetic resonance imaging were carried out. We measured the average ADC value of the renal lesions and normal kidneys. ADC maps from different b values were generated by a statistical package. Results: The ADC values of normal kidneys with three different motion-probing gradients(b=500, 800, 1000 sec/mm2) were 2.78±0.14×10-3mm2s-1, 2.45±0.13×10-3mm2s-1, 2.13±0.14×10-3mm2s-1, respectively. The ADC values of renal cell carcinoma with three different motion-probing gradients(b=500, 800, 1000 sec/mm2) were 1.63±0.14×10-3mm2s-1, 1.31±0.18×10-3mm2s-1, 1.07±0.15×10-3mm2s-1, respectively. Among the renal cell carcinoma, the ADC value of clear cell type were 1.67±0.09×10-3mm2s-1, 1.36±0.13×10-3mm2s-1, 1.15±0.14×10-3mm2s-1,respectively; the ADC values of granular cell type were 1.59±0.19×10-3mm2s-1, 1.25±0.22×10-3mm2s-1, 0.97±0.12×10-3mm2s-1, respectively. The ADC values of renal angiomyolipoma with three different motion-probing gradients(b=500, 800, 1000 sec/mm2) were 0.88±0.08×10-3mm2s-1, 0.63±0.07×10-3mm2s-1, 0.43±0.04×10-3mm2s-1, respectively. The ADC values of renal cystic lesions with three different motion-probing gradients(b=500, 800, 1000 sec/mm2) were 3.73±0.18×10-3mm2s-1, 3.44±0.13×10-3mm2s-1, 3.09±0.21×10-3mm2s-1, respectively. Statistically significant differences exists between the ADC values of normal kidney, renal carcinomas, renal angiomyolipomas and renal cysts when the b value is the same. Among the different cell types of renal carcinomas, the ADC value of granular cell carcinoma is lower than that of clear cell carcinomas. Conclusion: It is of benefit in diagnosing and distinguishing between benign and malignant renal tumors to know the ADC values in diffusion weighted magnetic resonance imaging. Furthermore, these values help to know the internal structure of the tumor and the tumor typel, which is helpful to the treatment and in predicting the patient’s prognosis.

Abstract:Objective: Studies have shown that TNF-αsecreted by tumor cells and macrophages infiltrated into the tumor microenvironment might promote the metastasis of a variety of malignant cancers, including breast cancer. The present study was designed to detect global metastasis-related gene expression changes of MCF-7 cells treated by low dose TNF-α and to further explore the mechanisms by which TNF-αcontributes to metastasis. Methods: MCF-7 cells were cultured and treated with low dose TNF-α(20 ng/ml). cDNA array analysis was applied to detect the metastasis related gene expressions. Results: A total of 36 gene expressions were significantly regulated by TNF-α. Functional analysis indicates that the altered genes belong to different functional group. Most of the genes changed may promote the metastasis of MCF-7 cells while the others may inhibit metastasis. The changes observed in gene expression following TNF-αwere somewhat time dependent. Conclusion: TNF-αcan enhance the invasive ability of MCF-7 cells, partly by regulating a series of metastasis related genes, and these genes may take part in every step of metastasis. Some of the genes deserve further study.

Abstract:Objective:To study the possibility of natural hydroxyapatite/chitosan composite on repairing bone defects. Methods:We developed a natural hydroxyapatite/chitosan composite that could be molded into any desired shape. The powder component consists of natural hydroxyapatite, which is epurated from bone of pigs. The liquid component consists of malic acid and chitosan. Operations were performed on the left tibias of 15 white rabbits to create two square bone defects. One of the defects was reconstructed with the composite, while the other was not repaired and used as a blank control. Three of the animals were killed at the end of 2 weeks, 4 weeks, 8 weeks, 12 weeks and 16 weeks respectively and implants were evaluated anatomically and histologically. Results:No apparent rejection reaction was found, except for a mild inflammatory infiltration observed 2 weeks after surgery. Fibrous tissue became thinner 2 ~8 weeks after surgery and bony connections were detected 12 weeks after surgery. The new bone was the same as the recipient bone by the 16th postoperative week. Conclusion:The hydroxyapatite/chitosan composite has good biocompatibility and osteoconduction. It is a potential repairing material for clinical application.

Abstract:Objective:Bruxism is the term used for teeth grinding or jaw clenching. An electronic monitor of bruxism was developed to evaluate bruxism duration and frequency. Methods: Ten cases were selected in the Department of Prosthodontics of Jiangsu Stomatological Hospital to evaluate the monitor. A stabilization occlusal splint was fabricated for each of the 10 cases. The vertical dimension for each splint was 0.5 mm lower than mandibular postural position. Some sensors had been prearranged at each splint which could transfer the variation of the biting force into electronic signals. The data of sleeping duration, grinding duration and grinding frequency were recorded with this new type of bruxism monitor, which had been specifically invented to study bruxism. Results:The data from 10 bruxism cases were collected and the results were considered reliable. Subjects nocturnal duration parameters did not change significantly from night-to-night. Conclusion:The bruxism monitor can automatically measure and record bruxism data using an occlusal splint. This device is valuable for diagnosis and evaluation of bruxism.

Abstract:Objective: The main purpose of this retrospective study is to evaluate clinical outcomes of Xive implants 3-year after insertion. Methods: 219 Xive implants were placed in 139 patients and their clinical effects were evaluated using the Albrektsson Standard. Results:The 3-year survival rate of the Xive implants is 97.26%(213/219). Eleven cases of immediate implants and 15 cases of implant insertion after maxillary sinus lifting all succeeded. The failure ratio is 6/219(2.74%) with failures resulting from excess bone loss in the implant region, peri-implant mucosal inflammation or continuous pain caused by some unknown reasons. Conclusion: Xive implants are clinically effective for the restoration of missing teeth. Immediate implant and implant following maxillary sinus lifting were also successful.

Abstract:Objective: To compare the effects of different methods of anesthesia and analgesia on the activities of phosphofructokinase(PFK), glucose-6-phosphate dehydrogenase(G-6PD) and aldose reductase(AR) in erythrocytes and levels of plasma glucose and stress hormones in patients undergoing esophagus surgery. Methods: Sixty-two patients scheduled for esophagus surgery were randomly divided into three groups: groupⅠ(n = 20) receiving only general anesthesia(GA) followed by intravenous patient controlled analgesia(PCA) with fentanyl 15μg/kg. The other two groups receiving both general anesthesia combined with thoracic epidural anesthesia (GEA) and either intravenous PCA with fentanyl 15μg/kg (groupⅡ, n = 21) or thoracic epidural analgesia(TEA) with 0.125% ropivacaine and 0.0002% fentanyl mixture(groupⅢ, n = 21) after the operation. Venous blood samples were collected for measurements of PFK, G-6PD and AR activities in erythrocytes and plasma glucose, cortisol, epinephrine and norepinephrine before induction (T1), 60 min following the incision (T2), 60 min(T3) after operation, on the 1st(T4) and 2nd postoperative day(T5). Results: The activities of PFK decreased(P < 0.01 or P = 0.004) and the activities of G-6PD and AR increased(P < 0. 01) in groupsⅠand Ⅱ on T4 compared with those on T1. Between the two groups, the activities of these enzymes in groupⅡchanged less than those of groupⅠ(P < 0.01 or P < 0.05). These enzymes activities changed slightly in group Ⅲ on T4(P > 0.05). There were significant differences between group Ⅲ and the other two groups(P < 0.01 or P < 0.05). The levels of plasma glucose increased significantly on T2(P < 0.01), reached peak values on T4(P < 0.01) and fell on T5 in the three groups. Compared to those of groupsⅠandⅡ, the values of plasma glucose in group Ⅲ were lower on T4 and T5(P < 0.05 or P < 0.01). The cortisol concentration in each group increased significantly at T2(P < 0.01 or P < 0.05), and remained elevated on T5(P < 0.01 or P < 0.05), while on T2 and T3 the cortisol levels of groupⅠwere higher than that of groupsⅡand Ⅲ(P < 0.05). The levels of group Ⅲwere lower than those of the other groups on T4 and T5(P < 0.01 or P < 0.05). The levels of epinephrine and norepinephrine were also significantly higher in group I than those of the other two groups on T2(P < 0.01 or P < 0.05), and their levels in groupⅠandⅡwere higher than that of group Ⅲ on T4. The patients of the three groups obtained satisfactory pain relief, with all Vidual Analogue Scale(VAS) scores less than 3. VAS scores of groupⅠwere much greater 4h after operation. Group Ⅲ VAS scores were the lowest 24h after operation. However, the number of times patients pressed the bolus switch was higher in groupⅡ(P < 0.01). Conclusion: Compared with GA and intravenous PCA, general anesthesia combined with thoracic epidural anesthesia and analgesia obtain better pain relief and could markedly alleviate the stress response and improve these erythrocyte glucose metabolism changes after esophagus surgery.

Abstract:Objective: To explore the measures and methods for improving distal phalanx replantation. Methods: One hundred and forty-eight cases with distal phalanx replantation from Sep 2004 to Sep 2007 were investigated. The indication for surgery and the operative program was determined by the trauma type and the degree of injury. As many microcirculation pathways as possible were reconstructed during the operation, and postoperative tissue decompression was also performed. Results: The survival rate of distal phalanx replantation was 89.9%(169/148). Conclusion: To improve the survival rate of distal phalanx replantation it is essential to reconstruct an effective local blood circulation, determine the degree of injury in cases with ecchymosis, and employ comprehensive postoperative measures.

Abstract:Objective: To explore the factors influencing insulin resistance in obese Chinese children. Methods: We randomly selected 53 children with uncomplicated obesity between 9 to14 years of age, and 29 normal healthy children, matched for age and sex. Anthropometric and plasma biochemical variables(including lipid profiles, glucose and insulin) were measured using standard methods. We calculated insulin resistance(IR) index using homeostasis model assessment(HOMA) methods and measured plasma high-sensitivity C-reactive protein(hs-CRP) levels using nephelometric methods. All statistical analyses were conducted using the statistical package SPSS. Results: Levels of fasting serum insulin, hs-CRP, total cholesterol(TC), low density lipoproteins cholesterol(LDL-C) and IR index were higher in obese children than in controls, while high-density lipoprotein cholesterol(HDL-C) values were lower in the obese children. There was no significant difference in levels of fasting blood glucose between the two groups. HOMA-IR was used as the dependent variable in multivariate regression analysis. Significant independent predictors for insulin resistance adjusted for waist/hip ratio, diastolic pressure (DBP), BMI, triglycerides and HDL-C level were waist circumference(WC), weight and systolic pressure(SBP). Conclusion: Waist circumference, weight and SBP are predictors of insulin resistance syndrome in Chinese adolescents

Abstract:Objective:To study the feasibility of the left radial approach for coronary angiography. Methods:195 patients diagnosed with coronary atherosclerotic heart disease were randomly divided for coronary angiography(CAG) into a left radial artery approach group(98 cases) and a right radial artery approach group(97 cases) from Jan 2006 to Dec 2006. Selective coronary angiographies were performed with 5F TIG catheters. The time of puncturing, duration under X-ray fluoroscopy and of the operation, successful rates of puncturing and coronary angiography were recorded. Results:There was no difference in the time of puncturing(2.25±1.58 min vs 2.19±1.62 min), duration under X-ray fluoroscopy(3.12±1.53 min vs 3.21±1.49 min) and the duration of the operation(12.87±2.52 min vs 12.98±2.85 min), nor in the success rates of puncturing(95.91% vs 95.87%) and coronary angiography(94.90% vs 94.85%). Conclusion: Coronary angiography can be accomplished via the left radial artery approach, indicating that this is a worthwhile clinical approach.

Abstract:Cetuximab is a new medication that has recently been approved for the treatment of advanced colorectal cancer. To date we have had little experience in using this targeted agent. Eleven patients in our hospital with advanced colorectal cancer were treated with cetuximab and chemotherapy. Based on the curative effect of this combination therapy, we have concluded that the following nursing practices make an important contribution to the patients’prognosis and wellbeing: to establish a good nurse-patient relationship, to increase patient understanding of the side effects, to standardize the medications, to observe and to deal with the side effects of the medications(for example skin reaction, neutropenia, and diarrhea), and to provide continuous mental health care support and education.

Abstract:Objective:To investigate the effect of insulin and blood glucose interference on interstitial cells of Cajal(ICC) in normal and diabetes mellitus(DM) mice stomach antrum. Methods:DM model was established by intraperitoneal injection of streptozotocin(STZ) in C57/BL6 mice. A week later,all the normal and diabetic mice were randomly divided into 4 groups,group A(normal control),B(normal mice administrated with high glucose),C(diabetic mice without interference),D(diabetic mice administrated with insulin). The blood glucose of all the mice was tested once a week. Six weeks later,the level of serum insulin was detected by ELISA,the amount of ICC was detected by Flow cytometry(FCM) and the ultrastructure of ICC in stomach antrum was detected by transmission electron microscope(TEM). Results:Compared with group A,there was a transient hyperglycemia in group B after injected glucose,and blood glucose in group C was significant inceased(P < 0.05). But there was no significant difference in blood glucose level between group A and D(P > 0.05). There was no significant difference in serum insulin level between group C or D,and no significant difference between group A and B(P > 0.05).While compared with group A and B,serum insulin level in group C or D was significantly deceased(P < 0.05). Compared with the group A,ICC quantity of the stomach centrum in B,C and D was significantly deceased(P < 0.05). There was no significant difference in ICC quantity between group C and D(P > 0.05). Compared with the group A,a significant ultrastructure change of ICC in group B,C and D was detected by TEM. The abnormal ultra-structure change of ICC in group B and C were most remarkable. Conclusions:There are quantitative decline and ultrastructure changes of ICC in DM mice stomach antrum. Both transient and persistence hyperglycemia may impair the ultrastructure of ICC and may cause ICC quantitative decline. Deficient of endogenous insulin may cause ICC quantitative decline,but not impair the ultrastructure of ICC directly. Add exogenous insulin can’t prevent from the quantity decline of ICC,but can prevent the ultrastructure change of ICC by controlling hyperglycemia.

Abstract:Objective:To construct a large human Na-觙ve Fab antibody library and to screen the anti-cMet phage antibody from the library. Methods:Human Fab genes from bone marrow lymphocytes of 20 healthy donors were amplified by PCR,and the products were ligated into the phagemid vector pComb3XSS to construct a human naive Fab library. Antibodies against c-Met were screened by biopanning with immobilized antigen. After six rounds of panning,sixty randomly selected clones were identified by phage ELISA to select specific ones with high affinity for c-Met. After analyzed by BstOI digest,the positive clone was used for soluble expression in Escherichia coli. and was characterized by ELISA. Results:A large human na-觙ve Fab phage-display library consisting of 1.2 × 109 clones was successfully constructed. From the enriched phage library,a Fab fragment designated AM2-26 with fine activity to c-Met was selected. DNA sequence analysis show the V genes were human variable region immunoglobulin sequences. AM2-26 was functionally expressed,which was identified by Western blot,and its specificity was confirmed by ELISA. Conclusion:The construction of the human phage antibody library and the preparation of the anti-cMet phage antibody provide a promising candidate for the research of antineoplastic agents.

Abstract:Objective:To investigate the expression of the recombinant plasmid of regulator of HIV-1 virion protein expression(Rev) in the eukaryotic cells,and to evaluate the effect on human herpesvirus 8(HHV-8) lytic cycle replication by Rev protein in BCBL-1 cells. Methods:After identification with enzyme digestion and nucleotide sequences analysis,the recombinant plasmid pRev-Flag was transiently transfected into BCBL-1 cells and NIH/3T3 cells. The expression of pRev-Flag mRNA and protein in both cells was detected by RT-PCR and Western blot,respectively. Subsequently,RNA was obtained from BCBL-1 cells transfected with pRev-Flag. RT-PCR was carried out to evaluate the expression of ORF26 mRNA(encoding the minor capsid protein) of HHV-8. Results:Nucleotide sequences analysis indicated that the cloned Rev sequence was 100% homology with Rev gene previously registered in GenBank. The specific band was detected at the expected place by both RT-PCR and Western blot. In addition,the expression of ORF26 mRNA in BCBL-1 cells transfected with pRev-Flag was weaker than the control. Conclusion:Rev gene was correctly expressed in BCBL-1 cells and NIH/3T3 cells,and Rev gene might inhibit KSHV lytic cycle replication.

Abstract:Objective:To study the effect of the centrifugal force on the constructing of tissue engineering nucleus pulposus. Methods:Nucleus pulposus were isolated by trypsin and type Ⅱ collagenase from rabbit thoracic and lumbar intervertebral discs.Cells were proliferated to the third generation and seeded into PLGA scaffolds(3 × 107/ml). Experimental groups were subjected to the centrifugal force(134.16g and 301.86 g of relative-centrifugal force,twice per day,for 20 min) while the control group was cultured under normal condition,each group containing 4 specimens. The complex of nucleus pulposus and scaffolds were observed after 4 weeks. The structure of nucleus pulposus and typeⅡ collagen secretion were assessed by HE,immunohistochemistrical stain and the scanning electron microscope. Results:After 4 weeks,all groups formed the similar of complex of nucleus pulposus and scaffolds. The tissue engineered nucleus pulposus under centrifugal force were morphologically thicker than that in control,the area of immunohistochemistrical stain was(14.36 ± 2.82)% in experimental group 1,(18.71 ± 2.14) in experimental group 2,(9.13 ± 1.58)% in control group respectively. There were statistical significance between experimental group and control group(P < 0.01),also between experimental group 1 and experimental group 2(P < 0.05). Conclusion:The centrifugal force promotes nucleus pulposus cells to secrete typeⅡ collagen and the centrifugal force might benefit the construction of tissue engineered nucleus pulposus.

Abstract:Objective:To establish the specific hybridoma cell lines that stably secreted monoclonal antibody(mAb) against human insulin-like growth factor-1 and identificate its properties. Methods:Balb/c mice were immunized with purified recombinant human IGF-1 protein. Splenocytes of the immunized mice were collected and fused with the mouse myeloma cell line Sp2/0 cells. The hybridoma cells that secreted human IGF-1 mAb were subcloned with limited dilution. The specificity of mAb was detected by Western blot. Indirect enzyme-linked immunosorbent assay(ELISA) was used to evaluate the affinity and titers of mAbs. The immunoglobulin (Ig) subtype of mAb was determined by test paper bar. Results:Two stably secreting anti-hIGF-1 mAb cell lines were obtained. Western blot analysis showed the mAb had specific combination with human IGF-1 which possessed biological activities. The titers of both mAb in ascites were 6 × 104 and their relative affinities were between 2.1 × 109/mol~7.8 × 109/mol. Two mAb cell lines can secret IgM subtype antibodies. Conclusion:Two hybridoma cell lines that secreted anti-hIGF-1 mAb were obtained,which will be useful for clinical application and fundamental basic research in hIGF-1.

Abstract:Objective:To establish the cultural system of hMSCs in vitro,investigate the changes of Bmi-1 gene expression in different passages,and explore the effect of Bmi-1 in hMSCs replicative senescence. Methods:hMSCs were isolated from the femur bone marrow using adherence method and cultured in L-DMEM with 10% fetal bovine serum(FBS). The 4 th passage hMSCs were picked up for analyzing expressions of cell surface phenotype,and the 1 st,3 rd,6 th,9 th,12 th,15 th and 20 th passages were examined of Bmi-1 gene expression by RT-PCR. Results:hMSCs were successfully isolated from the femur bone marrow which were positive for CD13,CD105,CD33 and negative for CD34,CD38,CD117,CD45. The SA-β-gal staining showed that the senescent cells were more suffused than the vigorous cells. The Bmi-1 mRNA was detected in hMSCs, and the gene expression decreased related to the cell passage. Conclusion:hMSCs that can be successfully isolated from femur bone marrow are easy to proliferate. The replicative senescence of hMSCs has correlation with the expression level of Bmi-1 gene.

Abstract:Objective:To explore the abundance of recombinant proteins expressed by the pET/E.coli system from different HCV gene regions. Methods:The four recombinant plasmids(pET-C\pET-NS5\pET-NS3-1\pET-NS3-6) were all transformed into the E.coli BL21(DE3) and screened by Kanamycin(50 -滋g/ml). After induced by isopropyl-β-D-thiogalactopyranoside(IPTG),the four expression products of fusion proteins were detected by 15% sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis(PAGE),and were purified by Ni-NTA Resin chromalography. The absorbances of the four recombinant proteins were measured by spectro photometer,and then the every protein concentration was calculated,respectively. Results:The four recombinant protein concentrations were 3.557(NS3 type 1),4.238(NS3 type 6),2.087(NS5) and 1.490 g/L(C),respectively. Conclusion:The result indicated that the abundance of four recombinant proteins was apparently different,derived by pET/E.coli system with different HCV gene fragments in the same condition.

Abstract:Objective:To evaluate the efficacy and safety of early postoperative transcatheter arterial chemoembolization in preventing recurrence of primary liver cancer and improving patients’ survival. Methods:Medline(1966-October 2007),Embase(1974-October 2007),the Cochrane Library(Issue 4,2007),CBMdisc(1978-October 2007),CNKI(1979-October 2007),Wanfang Database were searched without language limitation. All randomized controlled trials comparing hepatectomy alone with hepatectomy and early postoperative transcatheter arterial chemoembolization for primary liver cancer were identified and screened by two reviewers,and the methodological quality of the included randomized controlled trials were evaluated by Jadad scale. The Cochrane Collaboration’s RevMan4.2 software was used for analysis of the data extracted from the included randomized controlled trials. Results:Nine randomized controlled trials involving 586 patients were included with 6 trials published in Chinese and 3 in English. Meta-analysis of data extracted from the included randomized controlled trials showed that liver resection combined with early postoperative transcatheter arterial chemoembolization could significantly reduce recurrence rates compared with hepatectomy alone ,with the corresponding relative risk(95%CI) of 0.38(0.23~0.62)-0.61(0.41~0.90)-0.63(0.50~0.80)-0.74(0.60~0.90) at 6 month,1,3,5 year respectively; Meanwhile,early postoperative transcatheter arterial chemoembolization could also significantly improve overall survival,with the corresponding relative risk(95%CI) of 1.27(1.15~1.40)-1.52(1.25~1.85)-1.60(1.27~2.02)-1.78(1.09~2.90) at 1,2,3,5 year respectively. Side effects and severe adverse events related with postoperative transcatheter arterial chemoembolization were reported in 3 randomized controlled trials,mainly of liver function impairment,fever and gastrointestinal symptoms,and no death case related to treatment was reported. Conclusions:Based on the results of this Meta-analysis,early postoperative transcatheter arterial chemoembolization could reduce recurrence of primary liver cancer and improve the patients’ survival,especially in those with high recurrence risk factors,including large tumor volume with liver cirrhosis and tumor thrombosis. However,the methodological methods of most randomized controlled trials were in low quality,so more randomized controlled trials with high quality are still needed to assess and verify the efficacy and safety further.

Abstract:Objective:To explore the effection and mechanism of ischemia-reperfusion injury alleviation after performing liver transplantation on rats by pretreatment with ligustrazine. Methods:125 male SD rats were divided into 3 groups:sham operation group,control group(saline used as perfusate),experimental group(pretreatment with ligustrazine and saline used as perfusate). The rats in control group and experimental group were performed orthotopic liver transplantation. The levels of serum ALT,AST,TNF-α,IL-1-茁 and endotoxin were detected at 1,6,24,72 h after operation. Liver pathological changes was measured simultaneously. Results:the levels of serum ALT,AST,TNF-α,IL-1-茁 and endotoxin detected in experimental group were lower than those in control group,however higher than those in sham operation group(P < 0.05). Light microscopy result indicated that the inflammation and activation of Kuffer’s cells in experimental group were less severe than those in control group. Conclusion:Pretreatment with Ligustrazine could protect liver grafts against ischemia-reperfusion injury after liver transplantation in rats.

Abstract:Objective:To investigate the relationship between activation of NF-κB,expression of livin and survivin,two members of inhibitor of apoptosis protein(IAP) family,and drug resistance in gastric carcinoma. Methods:The inhibition of proliferation and apoptosis of gastric cancer cell SGC7901and SGC7901/VCR were detected by MTT and Flow cytometry. Activation of transcription of NF-κB was detected by ELISA. Protein expression of livin and survivin were assessed by Western blot analysis. Results:The inhibition of proliferation were reduced in SGC7901/VCR cells after been exposed to VCR,and the ratio of apoptosis was also decreased. We found that NF-κB activity was enhanced,and livin,survivin were overexpressed in SGC7901/VCR cells by ELISA and Western blot assay. Conclusion:The activation of NF-κB and upregulation of livin and survivin may be involved in the process of drug resistance in SGC7901/VCR cells . Upregulation of livin and survivin might relate to activation of NF-κB.

Abstract:Objective:This study was to determine the anti-proliferation effects of using rapamycin or adriamycin alone or combination of them on human breast cancer MCF-7 cells exposed to normal oxygen concentration or hypoxic enviroment in vitro. Methods:MTT assay and flow cytometry were used to examine the influence of the drugs on cell proliferation and cell cycle of MCF-7 cells. MCF-7 cells were exposed to normal or hypoxic enviroment and treated with rapamycin or adriamycin alone or co-treated with rapamycin and adriamycin. Western blot analysis was used to assess the expression level of HIF-1α and pAkt. Results:Opposite to adriamycin,the effect of rapamycin on inhibiting cell proliferation of MCF-7 cells was enhanced when exposed to hypoxia compared with the effect under normal oxygen environment. Regardless of oxygen concentrations,no synergistic interaction was observed when rapamycin at clinical normal drug level(10 ng/ml) combined with adriamycin on MCF-7 cells. The MCF-7 cells treated with rapamycin alone were arrested at G1,while co-treated with rapamycin and adriamycin were arrested at G2. Hypoxia enhanced the expression levels of HIF-1α and pAkt,while rapamycin at 10 ng/ml had no influence on the expression levels of HIF-1α and pAkt. Conclusions:Hypoxia increased the sensitivity of rapamycin on MCF-7 cells,which may be caused by increasing pAkt expression subsequent to hypoxia.

Abstract:Objective:To find potential molecular targets for breast cancer diagnostics by analyzing and comparing the proteomes expressed in human breast carcinoma,paracancerous tissues and normal tissues. Methods:The proteins expressed in these tissues were analyzed by the two-dimensional gel electrophoresis. The differentially expressed proteins in tumors were also analyzed by mass spectrometry(MS). Results:Protein extracts of individual sample in each type of tissues were separated on two-dimensional gels. There were twenty differentially expressed proteins in the tissues of breast cancer. Thirteen proteins were successfully identified by MS,in which eight proteins were overexpressed in tumors and the other five proteins were underexpressed. Conclusion:Significant discrepancies in protein expression exist among breast cancer tissues,paracancerous tissues and normal tissues. Some of the differentially expressed proteins found by the proteomic approach may be potential molecular targets for breast cancer diagnosis.

Abstract:Objective:To investigate the dynamic changes of vascular adventitia and smooth muscle cell(SMC) proliferation and collagen distribution in the vein graft restenosis model. To evaluate the effects of adventitia,cell proliferation and collagen distribution on vascular remodeling. Methods:The pig autogenous vein grafts restenosis model was constructed. Specimens were harvested at three time points that was 7th,30th and 45th days. According to the Masson staining slices,the vascular remodeling,proliferation index of adventitia cell and SMC,and collagen distribution were investigated by histomorphometrical and immunohistochemical approach. Results:①At 7thd after operation,the neointima formed and continuously thickened,whose thickness reached maximal at 45thd after operation. The thickness and cell density of adventitia increased gradually at 7thd,which reached peak at 30thd after operation. But in the 45thd group,they were smaller than those in the 30thd group. According to PCNA stain,the proliferation index of adventitia cell and SMC increased significantly at 7 d after operation,which reached peak at 30thd,whereas the expression of PCNA reverted toward the baseline at 45thd after operation. ②At 7thd after operation,the collagen of adventitia and neointima gradually increased,which of neointima at the 30th and 45thd group increased more significantly than the control and 7thd group(P < 0.05). The collagen of adventitia reached maximal at 30thd,but at 45thd,they were smaller than those at 30thd after operation(P < 0.05). ③ The luminal area and internal elastic lamina gradually decreased after operation,which in the 30thd and 45thd group reduced significantly compared with the control group and 7thd group(P < 0.05). But the residual restenosis rate changed inversely compared with the luminal area. Remodeling index and external elastic lamina slightly increased at the 7thd group,but reduced gradually after that,which in the 30thd and 45thd group decreased significantly(P < 0.05). Conclusion:The changes of vascular adventitia and smooth muscle cell proliferation,the thickness and fibration of adventitia and rearrangement of collagen are the important factors on intimal hyperplasia and vascular remodeling,which takes part in and accelerate the course of vein restenosis.

Abstract:Objective:To evaluated the effects of sildenafil on hemodynamics and cardioprotection in rheumatic heart disease patients with pulmonary hypertension undergoing valvular heart surgery during cardiopulmonary bypass(CPB). Methods:After pulmonary arterial catheter insertion,patients with mean pulmonary arterial pressures(MPAP) greater than 30 mmHg were eventually included in this study. Twenty-eight cases were randomly divided into the control group(n = 14),and the experimental group(n = 14). Each patient in the experimental group was given 100 mg SIL(orally by nasal gastric tube) after induction of anesthesia. In control group,patients were placebo-controlled. Hemodynamic variables were measured after induction of anesthesia,at 0,20,40,and 60 minutes after medication. Blood samples were taken to determine the serum concentrations of CK-MB and cTn-T before cardiopulmonary bypass,at 2,6 and 16 h after aortic open-clamp. Results:Compared with the control group,the mPAP and mPAP/PmSAP was significantly reduced(P < 0.01) in the experimental group. And the levels of CK-MB at 2h after aortic open-clamp and cTn-T at 16 h after aortic open-clamp in the experimental group were significantly lower than those in control group(P < 0.05). Conclusion:Sildenafil produced significant pulmonary vasodilatory effect in cardiac surgical patients with pulmonary hypertension and beneficial effects such as myocardial protection.

Abstract:Objective:To study the protective effect of prostaglandin E1(PGE1) on platelet during cardiopulmonary bypass(CPB).Methods:A total of 24 patients with rheumatic heart disease undergoing cardiac valve replacement were randomly divided into the experimental group(n = 12) and the control group(n = 12). The levels of TXB2 and 6-K-PGF1α in serum were measured at different intervals during and after CPB. The ultrastructure of platelets were also observed with transmission electron microscope. Results:The levels of TXB2 and 6-K-PGF1α during CPB were significantly higher than those before CPB in both groups(P < 0.01), and reached the highest at the end of CPB,then retrieved as before CPB. During CPB,the level of 6-K-PGF1α in the experimental group was significantly higher than that in the control group,while TXB2/6-K-PGF1α and the level of TXB2 were significantly lower than those in the control group(P < 0.01). Meanwhile,the number of unactivated platelets in the control group decreased more obviously than those in the experimental group during the CPB,while the number of activated ones increased(P < 0.01). Conclusion:PGE1 could effectively inhibit the activation of the platelets,protect their function and decrease their loss during the CPB,then reduce the coagulopathy resulted by CPB,reduce the amount of postoperative bleeding.

Abstract:Objective:To investigate the effect of peroxisome proliferator activated receptors-γ(PPAR-γ) agonist Rosiglitazone on the immune maturation of monocyte-derived dendritic cell(DC) induced by advanced glycosylation end products(AGEs). Methods:Monocytes were purified(over 98%) using Anti-CD14 microbeads. After cultured with DC Cellgro medium containing recombinated human granulocyte-macrophage colony stimulating factor(rhGM-CSF)(100 μg/L) and recombinated human interleukin-4(rhIL-4)(20 μg/L) for 5 days,monocytes were derived into immature DC. Rosiglitazone(50 μmol/L) was added to the medium for 24 hours. The advanced glycosylation end-human serum albumin(AGE-HSA)(200 μg/ml) was then added to the medium for another 48 hours. The expressions of(CD1a,CD80,CD86 and HLA-DR) were analyzed by FACS,and the cytokines secretions of culture supernatants(IL-10,IL-12,TNF-α and INF-γ) were measured by ELISA. Results:Rosiglitazone reduced AGE-HSA inducing expressions of CD1a,CD80,CD86 and HLA-DR,and also inhibited AGE-HSA inducing T-cell stimulatory activity. The cytokines secretions of DCs induced by AGE-HSA were significantly attenuated by Rosiglitazone. Conclusion:The study suggested that one of the anti-inflammatory mechanisms of PPAR-γ agonist Rosiglitazone should be mediated by inhibiting the AGEs induced maturation and immune function of DCs.

Abstract:Objective:To investigate the relationship between microalbuminuria and arterial stiffness index in diabetes patients. Methods:A total of 135 patients with diabetes were studied. Arterial Stiffness Index(ASI) which can reflect large arterial compliance was measured by CardioVisionMS-2000 arterial stiffness detector. Albumin/creatinine ratio(ACR) was determined. Results:ASI was higher in high ACR group than in normal ACR group. There were significant differences in age,SBP,PP,HDL-C,GFR,HbA1C,duration of diabetes between two groups(P < 0.05). There was a positive correlation between age,SBP,PP,LDL-C,ACR,HbA1C,duration of diabetes and ASI showed by single factor correlation analysis methods(P < 0.05) However there was a negative correlation between GFR and ASI(P < 0.05). PP,ACR,HbA1C,duration of diabetes were independent risk factors of ASI showed by multiple linear stepwise regression analysis method. Conclusion:The artery compliance as measured by ASI is increased in patients with type 2 diabetes and raised ACR.

Abstract:Objective:To explore the change of plasma angiotensinⅡ in patients with depression,and the relationship between angiotensinⅡand depressive symptoms. Methods:Thirty-one depressed patients diagnosed with Chinese Classification and Diagnostic Criteria for Mental Disorders criteria(3rd edition) were recruited and treated with citalopram for 6 weeks,and 30 healthy volunteers recruited as controls. The plasma angiotensinⅡ was measured with radioimmunoassay before and after treatment. The severity of symptom was evaluated with the Hamilton Rating Scale for Depression(HAMD) and Hamilton Rating Scale for Anxiety(HAMA). Results:The plasma angiotensinⅡ level was significantly higher in patients before treatment than that in controls[(436.75 ± 239.14)pg/ml vs(130.42 ± 47.85)pg/ml P = 0.000],and significantly decreased after treatment[(205.97 ± 155.00)pg/ml,P = 0.000]. There was significantly negative correlation between the reduction of plasma angiotensinⅡand the reduction rate of HAMD and HAMA score[HAMD(r = -0.457,P = 0.022);HAMA(r = -0.473,P = 0.017). Conclusion:It suggested that the angiotensinⅡmight be involved in pathophysiology of depression.

Abstract:Objectives:To measure fibrinolytic activity in the patients with tuberculous pleurisy and to evaluate the relationship between the effusion levels of these fibrinolytic components and the development of residual pleural thickening(RPT) in these patients. Methods:The plasma and effusion levels of PAI-1,D-D and t-PA were measured using an enzyme linked immunosorbent assay(ELISA)in 41 patients with TB,39 patients with lung cancer,11 patients with heart failure. pleural thickening was measured and defined as a pleural thickness of 10 mm found on chest radiographs at the completion of anti-TB chemotherapy in tuberculous pleurisy patients. Results:There was no significant difference in plasma PAI-1,t-PA and D-D levels among three diseases groups(P > 0.05). Tuberculosis exudates had significantly higher values for PAI-1 and lower values for t-PA than malignant exudates and transudate effusion caused by heart failure(P < 0.05). There was no significant difference in effusion D-D level in three groups(P > 0.05). Significantly higher levels of PAI-1 and significantly lower values of t-PA in effusions were found in patients with RPT than those in patients without RPT(P < 0.05). Conclusion:The release of PAI-1 increased while t-PA level decreased in TB pleural effusions. The imbalance of PAI-1 and t-PA may result in the development of pleural thickening in patients with TB pleurisy. Detection of pleural fluid D-dimer levels has no help in the differential diagnosis of an effusion.

Abstract:Objective:To investigate the effects of upper airway obstruction sites on polysomnographic parameters and clinical manifestations in patients with obstructive sleep apnea syndrome(OSAS). Methods:Thirty-six adult male OSAS patients with their AHI between 30~40 were selected to undergo overnight PSG test and simultaneous pressure measurement of upper airway and esophagus for identification of the lowest upper airway obstruction sites. Comparison was made among different obstruction sites(sites of soft palate,tongue-base and lower pharynx) for the following parameters:nocturnal mean arterial oxyhemoglobin saturation(meanSaO2) and minimal SpO2(miniSaO2),the longest obstructive apnea event time,rate of fall of SaO2(dSaO2/dt),arousal index; as well as some clinical manifestations:day-time epworth sleepiness scale,systolic blood pressure(SBp) at 9:00PM/SBp at 6:00AM,diastolic blood pressure(DBp) at 9:00PM/DBp at 6:00AM and incidences of hypertension. Results:There was no significant difference of mean SaO2,the longest obstructive apnea event time and SBp at 9:00PM/SBp at 6:00AM(P > 0.05) among three different lowest obstruction sites in OSAS patients. However,compared with the obstructive site at palate,the obstruction sites at tongue-base and lower pharynx displayed significant difference in the following PSG and clinical parameters:①lower miniSaO2(P < 0.05);②higher dSaO2/dt(P < 0.05); ③ higher arousal index and day-time epworth sleepiness scale(P < 0.01),lower DBp at 9:00PM/DBp at 6:00AM. (P < 0.05);④higher incidence of miniSaO2 < 80% and hypertension(all P < 0.01). Conclusion:In OSAS patients with similar AHI,the lowest upper airway obstruction site at tongue-base or lower pharnx may cause transcient hypoxiemia more efficiently and significantly as well as more severe clinical manifestations than the obstruction site at palate or higher.

Abstract:Objective:To study the relationship between insulin resistance and NAFLD. Methods:The age,BMI,WHR,TRIG,TC,and insulin sensitive index(ISI) were studied in NAFLD group and the control group. Results:The BMI,WHR,TRIG were higher in NAFLD than in the control,while the ISI was lower in NAFLD than in the control. The Logistic analysis show that WHR and ISI are important factors in the pathogenesis of NAFLD. Conclusion:WHR and ISI are the independent influential factors in the pathogenesis of NAFLD.

Abstract:Objective:To investigate the effects of anesthesia and postoperative analgesia with different methods on the differentiation of human helper T cells in the esophageal carcinoma patients undergoing thoracic surgery. Methods:Sixty esophageal carcinoma patients,classified as ASA physical status Ⅰ~Ⅱ,scheduled for elective thoracic surgery were randomly divided into three groups with 20 cases in each group:groupⅠreceived general anesthesia during thoracic surgery and patient-controlled intra-venous analgesia(PCIA) with fentanyl postoperatively;groupⅡunderwent general anesthesia plus thoracic epidural anesthesia(TEA) during thoracic surgery and received the same analgesia method as groupⅠpostoperatively; group Ⅲ were anesthetized as groupⅡ and received patient-controlled epidural analgesia(PCEA) with fentanyl and ropivacaine postoperatively. Venous blood samples for measurement of IFN-γ(as a marker of Thl cells) and IL-4(as a marker of Th2 cells)were taken before the induction of anesthesia and at set intervals until 48 h postoperatively. T helper cell subsets were quantified by using flow cytometry,and the ratio of Thl to Th2 cells was calculated. Results:The analgesial effects were similar among the groups. The Thl/Th2 ratio in all groups were decreased significantly after completion of surgery compared with baseline levels(P < 0.05). Group comparison,at T2 the Thl/Th2 ratio in GroupⅡ and Ⅲ higher than that in groupⅠ. In group Ⅲ the Thl/Th2 ratio at T3 and T4,were higher than those in groupⅡ. Conclusion:These findings show that general anesthesia combined with thoracic epidural anesthesia and PCEA may inhibit Th0 cells to differentiate into Th2 cells.