Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.

Objective To study the expression of epidermal growth factor receptor (EGFR) in cervical car-cinoma and cervical intraepithelial neoplasia(CIN),and to elucidate its relation with the genesis, infiltration, metas-tasis and prognosis of cervical carcinoma. Methods EGFR was determined by means of S-P immunohistochemistry in tissue of 100 cases of cervical carcinoma,60 cases of CIN and 40 cases of controls. Results The overexpression rates of EGFR were 0% (0/40), 51.67% (31/60),78.00% (78/100), respectively in normal cervical epithelium, CIN and cervical tumor tissues. The overexpression rate of EGFR was significantly higher in cervical tumor tissue than in control group(P<0.01). The overexpression of EGFR didn't demonstrate significant association with clinical staging, tumor size, pathological type, differentiation, cervical invasion depth, cervical canal invasion, lymphnode me-tastasis or the prognosis of cervical neoplasm (P>0.05). Conclusion Overexpression of EGFR is worsened with the severity of cervical lesion, suggesting that overexpression of EGFR is correlated with the genesis of cervical neo-plasms,which may be a valuable biological indicator of cervical carcinoma,but is not correlated with clinical patho-logical features and prognosis of cervical carcinoma.

Objective To explore a fast and precise registration algorithm for megavolt (MV) portal images(PIs) used for radiotherapy positioning verification, and find auto analysis method of set-up error using the computed image processing and mutual information comparison technology, which provide a basis for the development of automatic image guidance software. Methods MV PIs of patients undergoing radiotherapy were tested, pre-processed with noise reduction technique based on improved filtering algorithm and contrasted by gray-scale transforming using partial derivative threshold. The bone structures were then highlighted but soft tissues and the cavities were restrained simultaneously. Improved particle swarm optimization and powell hybrid algorithm were used to optimize and transform the mutual information based on wavelet multiresolution analysis when registering the Pls with digital reconstructed radiographs (DRRs) of treatment planning or X-ray simulation-film images(SIs). Application of the designed registration algorithm was verified and evaluated through simulated set-up shifts of head and neck phantom. Results The improved noise reduction algorithm satisfactorily met the requirements for contrast of bony structures in the MV PIs. The established mutual information registration method well behaved in both accuracy and speed of registration calculation. The processing of automatic registration took only 31.4 seconds averagely for the PIs and X-ray Sis of head-neck phantom. Mean errors of automatic registration of PIs and X-ray Sis in horizontal, vertical and rotational reduced by 62. 74% ,67. 32% and 66. 61% respectively compared with manual registration in the testing of 20-cases head and neck phantom. Conclusions A precise image registration algorithm and set-up error analysis method based on MV portal images is established, and it can meet the clinical application in registration accuracy and speed.

【Objective】 To study the expression of leptin mRNA in Chinese obese subjects using dot blot hybridization with a digoxigenin labele d probe. 【Methods】 11 Chinese nonobese subjects (bodymass index, BMI 21.0 kg/ m2±1.5 kg/m2) and 12 Chinese obese subjects(BMI 28.5 kg/m2±2.3 kg/m 2) parti cipated in the study. The human leptin cDNA probe was labeled wih digoxigenin(DI G) b y the random priming method. Expression of leptin mRNA in abdominal adipose tiss ue has been examined using dot blot hybridization with this probe. 【Results】 T he expression of leptin mRNA was significantly higher in obese subjects than in non-obese ones (312.8±108.9 vs 175.9±81.5, relative units, P<0.0 5), an d correlated with BMI (r=0.56, P<0.05). 【Conclusions】 Leptin mRNA le vel is high in obese subjects, and correlated with BMI.

AIM: To investigate the survival, migration and differentiation of human bone marrow mesenchymal stem cells (hMSC) in the middle cerebral artery occlusion (MACO) model and to observe the influence on motor function in the animal model. METHODS: hMSC with Hoeschst 33342 were injected into the animal model of MACO after Shenqiye induced for half an hour and their survival, migration, differentiation and the behavior changes in MACO rats were examined. RESULTS: hMSC had good homogeneousness and immunological reaction after implantation. The results showed that hMSC survived in rat brain for a long time over six weeks. As time going, hMSC were found in much more areas in the rat brain. Immunofluorescence staining suggested that implanted hMSC expressed human neuron specific enolase, neurofilament, and glial fibrillary acid protein. At the same time, improvements in abnormal behavior of MACO rats were observed. CONCLUSION: hMSC differentiate into neurons in the brain of rats, which means that hMSC is an ideal seed cells for the therapy of cerebral infarction.

The purpose of this study was to fabricate decelluarized valve scaffold modified with polyethylene glycol nanoparticles loaded with transforming growth factor-β1 (TGF-β1), by which to improve the extracellular matrix microenvironment for heart valve tissue engineering in vitro. Polyethylene glycol nanoparticles were obtained by an emulsion-crosslinking method, and their morphology was observed under a scanning electron microscope. Decelluarized valve scaffolds, prepared by using trypsinase and TritonX-100, were modified with nanoparticles by carbodiimide, and then TGF-β1 was loaded into them by adsorption. The TGF-β1 delivery of the fabricated scaffold was measured by asing enzyme-linked immunosorbent assay. Whether unseeded or reseeded with myofibroblast from rats, the morphologic, biochemical and biomechanical characteristics of hybrid scaffolds were tested and compared with decelluarized scaffolds under the same conditions. The enzyme-linked immunosorbent assay revealed a typical delivery of nanoparticles. The morphologic observations and biological data analysis indicated that fabricated scaffolds possessed advantageous biocompatibility and biomechanical property beyond decelluarized scaffolds. Altogether this study proved that it was feasible to fabricate the hybrid scaffold and effective to improve extracellular matrix microenvironment, which is beneficial for an application in heart valve tissue engineering.

Objective To verify the clinical effect of Qinghuang powder in chronic myelogenous leukemia(CML). Methods 86 CML patients treated with Qinghuang powder, in which 28 cases also partially received herbal medicine (activating blood circulation to dissipate blood stasis). Results 62 cases had complete remission (72.1%), 14 cases partial remission (16.3 %), advance 8 cases(9.3 %), inefficiency 2 cases (2.3 %), and the total efficiency was 97.7 %. The symptom were improved after a week when patients had taken medicines. 44 cases have hepatomegalia, among them 39 cases have diminished or became normal compared to untreated. 70 cases have splenoparectasis, among them 60 cases became normal, 9 cases diminished, and 1 case had no change after treatment. It took 15.5 days in average when spleen began to diminish, and took 62.9 days to become smallest. The WBC began to decrease at 10.4 day and took 54.8 days in average became normal. The major side effect was digestive tract symptom, followed by skin pigmentation and skin excessive cornification. It could be avoided by low dosage. Conclusion Qinghuang powder could not only induce CML to CR, but also improve the clinical symptom of CML and eliminate the infiltration of leukemia cells. It has little influence to Hb and Plt.

To develop a more efficient antithrombotic way after coronary artery bypass grafting (CABG), the anticoagulant effects were compared of human tissue factor pathway inhibitor (TFPI) gene transfection and aspirin oral administration (traditional method) on vein grafts. An eukaryotic expression plasmid pCMV-(Kozak) TFPI was prepared. Animal model of carotid artery bypass grafting was constructed. In operation, endothelial cells of vein grafts in TFPI group and empty plasmid control group were transfected with pCMV-(Kozak) TFPI and empty plasmid pCMV respectively, while no transfection was conducted in aspirin control group. After operation, aspirin (2 mg.kg(-1).(-1)) was administered (i.g.) in aspirin control group. Three days later, grafts (n=10) were harvested for RT-PCR, Western blotting and immunohistochemical analyses of exogenous gene expression and for pathological, scanning electron microscopic observation of thrombus. Thirty days later, the patency rates of remnant grafts (n=10) were recorded by vessel Doppler ultrasonography. Human TFPI gene products were detected in gene transferred vein grafts. Three days later, thrombi were found in 7 animals of aspirin control group and in 8 animals of empty plasmid control group, but in only 1 of TFPI group (P<0.01). Thirty days later, 5 grafts were occluded in empty plasmid control group, but none of grafts was occluded in the other groups (P<0.05). The endothelial surfaces of grafts in both of the control groups were covered with aggregated erythrocytes and platelets, and it were not seen in TFPI group. It was suggested that the anticoagulant effects on vein grafts of human TFPI gene transfection are better than those of aspirin.
Administration, Oral ; Anticoagulants/*metabolism ; Aspirin/*administration & dosage ; Aspirin/metabolism ; Coronary Artery Bypass ; Disease Models, Animal ; Lipoproteins/*metabolism ; Plasmids/metabolism ; Tissue Transplantation/*methods ; Transfection ; Ultrasonography, Doppler/methods ; Veins/*transplantation ; Venous Thrombosis/metabolism

Objective To study the biological characteristics of platelet-derived growth factor A and human beta defensin 2 (hPDGF-A/hBD2) gene-modified rat bone marrow mesenchymal stem cells (BMSCs). Methods By using liposome transfection technique, recombinant adenovirus vector expressing hPDGF-A/hBD2 (Adv-hPDGF-A-IRES-hBD2) labeled with GFP was transfected into 293T cells for virus packaging and amplification. BMSCs were isolated, cultured and infected by adenovirus-containing supernatant. The exogenous gene-modified BMSCs were comprehensively studied on their biological features, in terms of morphology, cell growth curve, cell cycle, and adipogenic, osteogenic and myogenic differentiation ability. Results hPDGF-A-IRES-hBD2 gene-modified BMSCs did not show obvious changes in cell viability, proliferation, cell cycle distribution or cell differentiation. Conclusion BMSCs were not only good carriers for exogenous hPDGF-A and hBD2 genes but also seed cells for cell therapy even after hPDGF-A/hBD2 modification.

Objective To investigate pathogens and drug resistance of lower respiratory tract infection(LRTI)in Intensive Care Unit(ICU).Methods Retrospective study of the clinical data,the distribution and the drug-sensitivity of pathogens of 220 cases with LRTI in ICU.Results Totally 280 strains of pathogens were identified by bacterial culturing.The ratio of G-bacteria to total pathogens isolated was 63.5%,of the G+ bacteria was 25.1%,and of the fungi was 11.4%.The main kinds of the G-bacteria were Klebsiella pneumoniae(17.1%),Pseudomonas aeruginosa(13.2%),Acinetobacter baumannii(12.5%),and Stenotrophomonas maltophilia(10.4%).Staphylococcus aureus(SA)(91.4%)was the most prominent in G+ bacteria,and MRSA was 98.4% in SA.The result of drug sensitive test in vitro showed the multiple drug fast rate of Pseudomonas aeruginosa was comparatively high,Stenotrophomonas maltophilia to Levofloxacin was low,Klebsiella pneumoniae and Acinetobacter baumannii were highly sensitive to carbapenems.The susceptibility rate of MRSA to vancomycin was 100%.Conclusion G-bacteria are the majority of the pathogens,isolated from patients with LRTI in ICU.Klebsiella pneumoniae,Pseudomonas aeruginosa,Acinetobacter baumannii,and Stenotrophomonas maltophilia are the chief G-pathogens.Except Stenotrophomonas maltophilia,imipenem and merpenem are relatively active against the G-bacilli.The proportion of MRSA and fungal infection is increasing.It is suggested that there be urgent need for surveillance of bacterial resistance and rational use of antimicrobial agents during clinical therapy.