Adenovirus as gene transfer vector is characterized by high transfection efficiency,high titer,good stability in target cells and low pathogenicity,and has been widely used in gene therapy.Compared to first-and second-generation adenovirus vectors,the third-generation adenovirus vectors have unique advantages for gene therapy because of their low immunogenicity and high capacity.However,the clinical application of the third-generation adenovirus vectors is hampered by the existance of helper contamination and difficulties in their large-scale production.

Trypsin was purified from crude material of pig pancreas by AOT/isooctane reversed micellar system.The influence of the main operating parameters such as ethanol concentration in forward and backward extraction,pH,KCl and AOT concentration,temperature were investigated.Forward and backward extraction recovery of trypsin reached almost 90% and neared 100%,respectively.Finally,about 88% of total yield was obtained,and the specific activity of trypsin was increased to over 1800U/mg with purification factor of 5 folds more.In AOT-isooctane reverse micellar extraction system,denaturation or precipitation of proteins always occured due to strong electrostatic interaction between AOT-proteins molecules.It had been resolved by adding ethanol into reverse micellar system,and no denaturation was observed.Otherwise,the phase separation time was shortened significantly because of ethanol added.It was only 10 minutes or less to reach phases separation after forward and backward extraction.If this method can be applied in industry,efficiency will be greatly improved.

Objective:To observe the histological character of distraction osteogenesis(DO) in the mandible of goat using an intraoral tooth-supported distraction device. Methods:8 goats (7-9 months of age) were divided into 4 groups with 2 goats per group. After a bilateral mandibular corticectomy between the second and third premolars, self-produced intraoral tooth-supported distraction devices were cemented. 5 days after operation,the distraction devices were begun to be activated 0.25 mm every 12 hours to advance the anterior segment 8 mm in 16 days. The animals of 32 days group were injected with tetracycline fluorescence labeing at 10 ml/kg once perweek from the beginning of DO.The animals were sacrificed on day 8,16,32 and 48 separately post activation, and tetracycline fluorescence labeling,HE staining and Mallory's 3-color staining were performed. Results:Tetracycline fluorescence labeling showed that there was green-yellow fluorescence labeling new bone within the distraction gap; HE and Mallory's trichrome staining showed that new bone gradually formed from bilateral sides, gradually blending and remodeling. The fiber bundles and newly formed trabeculas and various cells in early phrases showed to be orientation-directed along the axis of distraction force. Conclusion: New bone can be formed and regenerated along the axis of distraction force within the distraction gap.

Adult ; Humans ; Laryngeal Neoplasms ; Male ; Neurofibroma ; Vocal Cords ; pathology

Objective To discuss color Doppler uhrasonographic (CDU) grading of varicocele (VC) in predicting the changes of the fertility index (FI) after varicocelectomy.Methods One hundred and fifty-one patients with VC were diagnosed and graded by CDU.All patients were grouped by the regurgitant volume and the maximum diameter of pampiniform plexus veins(PPV).The relation between the regurgitant volume and the maximum diameter of PPV and the changes of the FI after varicocelectomy was analyzed.Results The pathological regurgitation could be detected when the PPV diameter > 1.5 mm.The positive correlation was observed between the regurgitant volume and the diameter of PPV (r = 0.89, P <0.01).The pre- and postoperative FIs were extremely significant difference in regurgitation grade Ⅱ and regurgitation grade Ⅲ (P < 0.01) but not significant difference in regurgitation grade Ⅰ (P>0.05).The pre- and postoperative FIs were no significant difference in the group with the PPV diameter ≤ 2.0 mm (P > 0.05), but statistically significant difference in the groups with the diameters 2.1-2.5 mm and 2.6-3.0 mm (0.01 < P <0.05).FIs in the group with the diameter > 3.0 mm exhibited extremely significant difference after varicocelectomy (P < 0.01).Conclusions CDU grading of VC could be used to predict the changes of seminal quality after varicocelectomy.Varicocelectomy isn't necessary to the patients with VC that the PPV diameter ≤ 2.0 mm or pathological regurgitation being detected only on Valsalva maneuver.

BACKGROUND:Ricin possesses strong cytotoxicity but lacks of specificity.Severe adverse reactions can be caused if ricin is systemically administered.Interstitial chemotherapy with ricin coated by a sustained-release carrier can enhance local therapeutic effects of tumor,avoiding systemic-toxic occurrence.OBJECTIVE:To prepare injectable and biodegradable thermosensitive poly(D,L-lactide-co-glycolide)(PLGA)-polyethylene glycol(PEG)-PLGA copolymer-based hydrogels and to observe the in vitro drug delivery of the PLGA-PEO-PLGA copolymer hydrogel as a sustained-release carrier of ricin.DESIGN,TIME AND SETTING:A controlled observation was performed at the Center for Medical Bioengineering.Fujian Medical University between August and December 2007.MATERlALS:Ricin with a purity of 95%was purified by Fuiian Institute of Ultrasonic Medicine.DL-lactide and L-glycolide were provided by Glycolide Lactide Co.,Ltd..China.METHODS:PLGA-PEG-PLGA-copolymer was synthesized by ring-opening polymerization.MAIN OUTCOME MEASURES:Phase transition temperature was determined by test tube turnover.relativemolecular mass by high-performance gel permeation chromatography,the proportion of block copolymer by nuclear magnetic resonance spectrometry.the in vitro drug delivery of ricin coated by PLGA-PEG-PLGA copolymcr hydrogelby enzyme linked immunosorbent assay.the structural integrity of released ricin by high-performance liquid chromatography,the cytotoxicity of released ricin by methyl thiazolyl tetrazolium assay.RESULTS:The synthesized copolymer PLGA-PEG-PLGA exhibited reverse.thermosensitivity.The phase transition temperature of the copolymer was 26℃at 230 g/L.The weight-average molecular weight and number-average molecular weight of this copolymer was 8 027 and 6 643,respectively,and the multiple-dispersion toefficient was 1.17.The block ratio for PEG and PLGA was 3:2.Drug-loaded PLGA-PEG-PLGA hydrogel could release ricin in vitro for a total of 18 successive days and simultaneously well maintain cytotoxicity and the structural stability of ricin.CONCLUSION:PLGA-PEG-PLGA copolymer is an ideal carrier for sustained delivery of ricin

Objective To investigate the value of ultrasonography in diagnosis and preoperative TNM staging for neuroendocrine breast cancer(NEBC).Methods The ultrasonic characteristics of 12 NEBC and 104 invasive ductal carcinoma(IDC) confirmed by pathology,as well as the metastatic lymph nodes were analyzed retrospectively.The TNM-staging for NEBC by preoperative ultrasonography was evaluated according to pathology.And the relation between lesions size and Ki-67 was then analyzad.Results NEBC lesions mainly showed solid and heterogeneous hypoechoic(66.67％),irregular shape(66.67％),clear border (58.33 ％),posterior enhancement(58.33 ％) and internal blood flow(grade 0-Ⅰ) (66.67 ％).The detection rate of unclear border,irregular edge,hyperechoic halos and uneven internal echo was lower than IDC(P ＜ 0.05).Meanwhile,NEBC had a lower detection rate of calcification and internal blood flow(grade Ⅱ-Ⅲ)than IDC (P ＞0.05).And the detection rate of more round or oval lesions were higher in NEBC than that in IDC (P =0.001).According to pathology,the ultrasonic coincidence rate for T staging of NEBC and pathology was 75％,and for T1-staging was 100％,for T2-staging was 80％.The sensitivity,specificity,positive predictive value and negative predictive value of ultrasonic diagnosis on lymph node metastases was 66.67％,88.89％,66.67％ and 88.89％,respectively.There was a positive correlation between focal maximum diameter with Ki-67 (r=0.026,P=0.004).Conclusions It shows certain value of ultrasonography in diagnosis and preoperative TNM staging for NEBC.

ObjectiveTo assess the clinical value of ultrasound-guided percutaneous ethanol injection (PEI) in the treatment of hepatocellular carcinoma (HCC) combined with portal vein tumor thrombosis (PVTT).MethodsNinety HCC patients with different type PVTT underwent ultrasound-guided PEI once or twice a week.The follow-up lasted 12 to 60 months,and the 1-,2-,3-,4- and 5-year survival rates were calculated.ResultsThe median survival time of HCC combined with PVTT patients treated with PEI was 12 months,and the 1-,2-,3-,4- and 5-year survival rates was 52.22％,23.33％,15.56％,12.22％ and 10.00％,respectively.The survival rate correlated significantly to the type of PVTT.The lower the type of PVTT was,the longer the patients lived.ConclusionUltrasound-guided PEI is a safe and effective method for the treatment of HCC combined with PVTT,and can prolong the survival time of HCC patients with low type PVTT.

Objective To examine the effects of IL-10 on cardiac fibroblasts ( CFBs) proliferation and phenotype transformation to myofibroblasts (MyoFbs) induced by transforming growth factor-β1 (TGF-β1);and to investigate the regulating pathways .Methods Cardiac fibroblasts were isolated from cardiac ventricles of neonatal SD rats . The passage 2~4 were used and divided into the following groups for treatment:1) control group, 2) IL-10 reac-tion group, 3) TGF-β1 reaction group, and 4) IL-10 plus TGF-β1 reaction group (TGF-β1 treatment followed with IL-10 pretreatment ) .Cells proliferation was assessed by MTT assay and immunocytochemistry staining for prolifera-ting cell nuclear antigen (PCNA);the phenotype transformation into MyoFbs was assessed by immunocytochemistry of α-smooth muscle actin (α-SMA);extracellular signal related kinase ( ERK1/2) and P38 kinase pathways were assessed by western-blot.Results TGF-β1 (10 μg/L) treatment boosted the proliferation and the expression ofα-SMA significantly (P<0.01), while IL-10 (10, 50 or 100 μg/L) plus TGF-β1 co-treatment induced lower cell proliferation and expression of α-SMA than treating with TGF-β1 alone ( P<0.05 ) , with the inhibitory effect of IL-10 being concentration dependent .TGF-β1 could significantly stimulate the ERK 1/2 and P38 kinase phospho-rylation ( P<0.01 ) , however IL-10 (100 μg/L) plus TGF-β1 co-treatment failed to down-regulated the phospho-rylation of ERK1/2 and P38 kinase compared with TGF-β1 alone ( ERK1/2:P<0.05;P38:P<0.01 ) .Conclu-sions IL-10 can attenuate TGF-β1-induced CFBs proliferation and phenotype transformation to MyoFbs .The in-hibitory effects may explained by a mechanism of inhibiting the activation of ERK 1/2 and P38 kinase .

Objective To estimate the degree of oxidative stress and atherosclerosis praecox in patients with systemic lupus erythematosus (SLE), and emphasize on exploring AOPPs' clinical significance in premature atherosclerosis in SLE. Methods The levels of AOPPs, Hey, MDA, SOD, baPMV and CIMT were detected by ELISA and spectropholometry in 44 non-menopausal female SLE patients and 31 healthy middle-aged women respectively, and baPWV. The results were compared with AOPPs of the two groups. Then each group was stratified based on disease duration (≥5 years or <5 years) and the disease activity(active and inactive) in SLE patients. The patients' TC, TG, LDL were analyzed. T test, t' test and Pearson correla-tion were selected. Results The levels of AOPPs, Hcy, MDA in SLE patients were significantly higher than those of the healthy controls (P<0.05). The activities of SOD were lower than controls (P<0.05). The levels of AOPPs, Hcy, MDA, SOD had statistical significance between SLE patients disease duration ≥5 years or <5 years, active or inactive groups. There were two cases with CAP in patients with SLE (more than 5 years disease duration),while there wasnone in healthy controls. The levels of baPWV and CIMT in patients with SLE were higher than healthy controls (P<0.05), and had statistical significance in SLE patients disease duration more than or less than 5 years (P<0.05). The oxidative stress targets (AOPPs, Hey, MDA, SOD) had significant correlation with the level of baPWV and CIMT (P<0.01). The level of serum AOPPs had significant positive correlation with the levels of Hcy, TC, TG and LDL (P<0.01～0.05). Conclusion SLE patients have increased oxidative stress , and significantly higher prevalence of atherosclerosis than healthy controls. The disease duration and oxidative stress play important roles in the duration of atherosclerosis. AOPPs probably involves in the accelerated atherosclerosis of SLE patients. It may be a predictor for SLE complicated with atherosclerosis praecox.