Objective:To construct a monoclone cell line expressing M-HBsAg.Methods:Transfected SP2/0 with pRc/CMV-S 2S by Calcium Phosphate Method,selected the cells by G418,detected HBsAg by Dot-blot、ELISA and Western-blot hybridization.Results:By selection of G418,G418 resistent cells were obtained;tests of Western-blot hybridization and ELISA demonstrated their expression of aimed protein.Conclusion:Monoclone Cell Lines of SP2/0-S 2S were obtained.

Objective To compare the diagnostic performance of prospective electrocardiogram (ECG)-gating 320-detectorcomputed tomography coronary angiography (CTCA) versus retrospective ECG-gating CTCA. Methods 500consecutive patients suspected coronary artery disease with heart rate of less than 65 bpm were performed coronary computed tomography angiography (CCTA) with prospective (group P)and retrospective (group R) ECG gating in turn. The image quality was divided into four levels. Success rates, effective radiation dose,image quality and diagnosis were evaluated. Results Success rates of examination in group P and R are100%.Mean patient radiation dose was significantly different in group P (3.28±1mSv) and R(14.36 ±2.3 mSv).131 and 142 of 3750 segments (250 patients x 15 segments per patient) were too small(1.5mm)tobe characterized in group P and R. 96. 51% (3619/3750) and 96.12% (3608/3750) characterized segments were not significantly different in group P and R. 95.09% segments received a score of 4 and 1.09% segments received a score of 3 in group P, while 95.23% and 0.74% in group R.The cause of 2 and 3 points in the two groups was motion artifact. Stair-step artifacts and images scored as 1were not found.Compared with CAG,the sensitivity,specificity,false positive andfalse negative value in group P(86. 49% ,98. 70% ,88. 89% ,98.38%) and R(83. 87% ,98.74% ,89. 65%,97.93%) were not significantly different. Conclusion Although the patient radiation exposure was significantly lower, the diagnostic performance of prospective ECG-gating 320-detector CTCA was comparable with that of retrospective ECG-gating 320-detector CTCA on patients with stable heart rates less than 65 bpm.

Objective Using CT perfusion (CTP) technique,to investigate the graft perfusion changes in patients with hepatic artery stenosis (HAS) with or without ischemic-type biliary lesions (ITBL) after orthotopic liver transplantation (OLT).Methods Thirteen recipients with HAS received CTP scan of the liver,including 8 with ITBL and 5 without ITBL.For all patients,the diagnosis of HAS was made by CTA,and the diagnosis of ITBL by percutaneous transhepatic cholangiography.CT perfusion indices were obtained,including hepatic artery perfusion (HAP),portal vein perfusion (PVP),total liver perfusion (TLP) and hepatic perfusion index (HPI).Results Of the 13 patients with HAS,mean HAP in patients with and without ITBL was 59.8 and 35.1 ml·min-1 ·100 ml-1 (P =0.021,two-tailed paired Student t test) ; mean PVP was 125.4 and 166.2 ml·min-1·100 m1-1 (P =0.016) ; mean TLP was 185.2 and 201.3 ml· min-1 · 100 ml-1 (P =0.306) ; and mean HPI was 33.6 and 18.2 (P =0.005),respectively.Conclusion Using CTP technique,liver perfusion changes were reflected by measuring CTP indices noninvasively.Compared to those without ITBL in this study,HAP and HPI in patients with ITBL were higher and PVP was lower,which may be contributed to biliary inflammation.

ObjectiveTo investigate the outcome and risk factors for kidney involvement by analyzing 64 patients with anti-neutrophil cytoplasmic antibody(ANCA)-associated vasculitis.MethodsData analyzed including the demographic information,survival status,renal survival status and laboratory parameters such asserum albumin level,serum creatinine level,urinary protein excretion level,hematuria,high sensitivity C-reactive protein(CRP),ANCA titer,and the Birmingham vasculitis activity score (BVAS).Logistic regression analysis,Cox regression analysis and ROC curve were used to evaluate the risk factors of patients with renal involvement and all-event survival.ResultsTotally 64 patients were enrolled [24 females with the average age of (59.9±2.0) years] and followed up for a median of (38±16) months.The morality rate was 14％,and the prevalence of end stage renal disease was 39％.Compared with those who had better outcomes,patients who died or with end stage renal disease had higher serum creatinine level [ (624±246),(245±127 ) μ mol/L,respectively,t=7.17,P＝0.005 ] and erythrocyte sediment rate [ (112±24),(76±48) mm/1 h,respectively,t=3.74,P＜0.01 ],but lower serum albumin level [(294±31 ),(316±42) g/L,respectively,t=-2.27,P=0.01 ] and hemoglobin level [ (79±13),(99±33) g/L,respectively,t=-3.23,P＜0.01 ] at baseline.Logistic regression analysis found that serum creatinine level and erythrocyte sediment rate at baseline were associated with poor outcome and Cox regression analysis further confirmed this result[Scrβ=1.004,95％CI1.002～1.006,P＜0.01; ESR β=l.018,95％CI 1.000～1.037,P=0.046].ROC curve analysis showed that serum creatinine and erythrocyte sediment rate were predictors for AAV patients' prognosis and their AUC were 0.95 and 0.80,the sensitivity of these parameters was both 94％,and the specificity was 93％ and 70％respectively.ConclusionThe intensity of initial treatment should be based on disease severity and activity in order to improve the prognosis of those with ANCA-associated vasculitis with renal involvement.Increased serum creatinine and erythrocyte sediment rate may serve as predictors for poor prognosis in this patient cohort.

OBJECTIVE: To assess the diagnostic value of CT peritoneography for peritoneal complications of continuous ambulatory peritoneal dialysis (CAPD). METHODS: CT peritoneography in 11 CAPD patients with clinically suspected dialysis-related complications was prospectively studied. The CAPD patients were all treated in Renji Hospital from 2005 to 2007. CT images were reviewed according to the evidence of peritoneal leaks, hernias, loculate pleural fluid collections, and adhesions. RESULTS: Abnormal findings were found in 9 of 11 CAPD cases including inguinal hernias (3 cases), umbilical hernia (1 case), hydrocele (1 case), leaks in catheter tunnel (2 cases), and peritoneal adhesions (2 cases). Parts of them were confirmed by surgical operation. CONCLUSION: CT peritoneography is useful for the evaluation of complications related to CAPD, and it offers excellent tissue contrast and multiplanar imaging for assessment of the complications.

OBJECTIVETo explore whether ultrasound-mediated microbubble destruction can enhance the expression efficiency of plasmid pIRES-rhBMP2-EGFP for bone morphogenetic protein-2(BMP-2) in mice skeletal muscle.

METHODSTwenty four male BALB/c mice were divided into four groups. The naked plasmid was injected into the pretibial muscle or the quadriceps muscle (group A and group C) without ultrasound-mediated microbubble destruction method. Micobubbles with plasmid were injected into the pretibial muscle or the quadriceps muscle (group B and group D) with destructing microbubbles by ultrasound immediately. Twelve mice (group A and group B, 30 microg plasmid injected) were killed after 7 days and the tissue samples of the pretibial muscle were obtained to observe the expression of enhanced green fluorescence protein (EGFP) by inverted fluorescence microscope, gene transfection efficiencies were quantified by counting EGFP positive fibers on mice skeletal muscle. After 14 days, the other twelve mice (group C and group D, 100 microg plasmid injected) were killed and immunnohistochemical technique was applied to detect the rhBMP-2 gene expression.

RESULTSThe percentage of GFP-positive fibers was significantly lower in the group A than that in the group B. After 14 days, expression of rhBMP-2 was detected in cells and interstitial spaces in the group C and group D, and expression efficiency of rhBMP-2 in the group D was significantly higher than that in the group C.

CONCLUSIONUltrasound-mediated microbubble destruction could enhance the transfection and expression efficiency of rhBMP-2 gene in skeletal muscle of mouse in vivo. It is a new gene therapy method for periodontal regeneration.

Animals ; Bone Morphogenetic Proteins ; Genetic Therapy ; Green Fluorescent Proteins ; Male ; Mice ; Mice, Inbred BALB C ; Microbubbles ; Muscle, Skeletal ; Plasmids ; Transfection ; Ultrasonics

Sulfur fumigation (SF) is a universal phenomenon in primary processing of Traditional Chinese Medicine (TCM) in modern times. In the process, fumigation, sulfur or both of them act on the TCMs. Some active components of TCMs change quantitatively or qualitatively during the processing. At the same time, the sulfur dioxide and heavy metal would remain and cause a serious influence on quality and future development of TCM. This article reviews the chemical compositions change after SF to study the change law and their influence on quality. This article provide references for SF in TCMs' processing for a better and safer quality.
Drug Contamination ; Fumigation ; methods ; Medicine, Chinese Traditional ; methods ; Quality Control ; Sulfur ; chemistry ; Technology, Pharmaceutical ; methods

Infestation, moldy and other phenomenon in the processing and storage of Chinese herbal medicines is a problem that faced in the production of Chinese traditional medicine. The low productivity of traditional processing methods can not guarantee the quality of Chinese herbal medicines. Sulfur fumigation is the first choice of grassroots to process the Chinese herbal medicine with its low cost and easy operation. Sulfur fumigation can solve some problems in the processing and storage of Chinese herbal medicines, but modern pharmacological studies show that long-term use of Chinese traditional medicine which is fumigated by sulfur can cause some serious harm to human liver, kidney and other organs. This paper conducts a review about the application history of sulfur fumigation, its influence to the quality of Chinese herbal medicines as well as domestic and foreign limits to sulfur quantity, and a brief introduction of the status of modern processing technologies in the processing of food and some Chinese herbal medicines, the problems ex- isting in the Chinese herbal medicines processing, which can provide a reference basis for the further research, development and application of investigating alternative technologies of sulfur fumigation.
Fumigation ; adverse effects ; methods ; Medicine, Chinese Traditional ; methods ; Quality Control ; Social Control, Formal ; Sulfur ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate the transfection efficiency of the recombinant human bone morphogenetic protein-2 (hBMP-2) gene in targeted cells by ultrasound-mediated microbubble destruction.

METHODSNIH3T3 cells' anabiosis was completed and went down to the 3rd or 4th generation, and cultured in 6 well plates. The cells were divided into 2 groups: Plasmid DNA and Lipofectamine 2000 group (liposome group), plasmid DNA and ultrasound and microbubble group (ultrasound-mediated microbubble destruction group). Plasmid DNA was transfected into cells with liposome or ultrasound and microbubble. 24-48 hours later, the transfection efficiency and the concentrations of hBMP-2 were measured with fluoresence microscope and enzyme-linked immunosorbent assay (ELISA) respectively. The data were analyzed by curve fitting and t-test of SPSS 11.5.

RESULTSThe transfection efficiency rate was (7.30 +/- 1.58)% in liposome group, compared with (11.77 +/- 3.16)% in ultrasound-mediated microbubble destruction group (P< 0.05). The concentration of hBMP-2 after transfection was (1164.35 +/- 724.67) pg/mL in liposome group, versus (2932.70 +/- 656.27) pg/mL in ultrasound-mediated microbubble destruction group (P<0.05).

CONCLUSIONUltrasound-mediated microbubble destruction could significantly improve the transfection efficiency and expression of hBMP-2 gene in NIH3T3 cells. It may provide a new and effective gene delivery system for gene therapy in periodontal regeneration.

Animals ; Bone Morphogenetic Protein 2 ; Genetic Therapy ; Humans ; In Vitro Techniques ; Mice ; Microbubbles ; NIH 3T3 Cells ; Plasmids ; Recombinant Proteins ; Transfection ; Transforming Growth Factor beta ; Ultrasonics

OBJECTIVETo observe and compare the effects of 17beta-estradiol (EST) on the phasic and tonic contractile activities of the uterine smooth muscles of SD rats in vitro.

METHODSDifferent concentrations of 17beta-estradiol were added into the perfusion muscular sockets containing uterine smooth muscles of SD rats, and the activities of muscle contraction were recorded at the same time.

RESULTS17beta-estradiol had obvious depression effects on spontaneous rhythmic contraction of the uterine smooth muscles in a concentration-dependent manner, it could considerably decrease muscular tension, the mean amplitudes and frequencies of contractile waves (P < 0.01); it could also suppress the uterine contraction stimulated by KCl, CaCl2 or prostaglandin F2alpha (PGF2alpha). Based on the contraction of uterine smooth muscle stimulated by KCl, IC50 was 7.278 micromol/L and pD2 was -0.862 when calculated by linear regression method. 17beta-estradiol could also inhibit the maximal CaC12 contraction of uterine smooth muscle in the Ca2+ free Krebs solution, which the ECQ was 1.422 x 10(-3) mol/L, pD2 was 2.847 (control), but the E50 was 3.028 x 10(-3) mol/L, p2 was 2.519 (added with EST) when calculated by linear regression method.

CONCLUSIONThe depression effects of 17beta-estradiol on the spontaneous rhythmic contraction and activated contraction of the uterine smooth muscles of SD rats could be mediated through the blockage of C2+ influx through potential-dependent Ca2+ channels of plasma membrane.

Animals ; Estradiol ; pharmacology ; Female ; Muscle, Smooth ; drug effects ; Myometrium ; drug effects ; Rats ; Rats, Sprague-Dawley ; Uterine Contraction ; drug effects