To better understand the cleavage efficiency of muhiribozyme system on its RNA substrate in the presence and absence of divalent magnesium and monovalent sodium ions.we constructed pGEM-Coat'A,pGEM-Coat'A196Rz plasmids and pGEM-MDRl target plasmid.They were applied to transcribe RNAs with SP6/T7 transcription kit.Cleavage reactions were carried out in cell-free system and reaction products were analyzed by electrophoresis on 6% denaturing polyacrylamide gels in TBS buffer.The gels were dried and exposed to X-ray films for autoradiography.The Image J software was employed to analyze the dried gels.The results indicated that the cleavage efficiency of the muhiribozyme was dependent on the concentration of divalent Mg2+.The cleavage products increased with the concentrations of divalent Mg2+ and were Mg2+ concentration and time dependent.No cleavage product was obtained in the presence of lower than 200 mmol/L Na+ alone.On the contrary,monovalent Na+ inhibited the Mg2+ -induced cleavage reaction in Na+ and Mg2+ coexistance.The cleavage rate was significantly lower than that observed with divalent Mg2+ alone.These results suggested that divalent Mg2+ was required for muhiribozyme on substrate cleavage reaction in the physical condition,whereas monovalent Na+ was not.

Objective To compare the sonographic and pathologic features of calcified and non-calcified ductal carcinoma in situ DCIS Methods A total of 83 lesions in 82 consecutive patients with pathologically confirmed pure DCIS were recruited One patient had bilateral lesions All lesions were divided into calcified DCIS and non-calcified DCIS according to the presence of calcifications on mammography Their sonographic features and pathologic reports for all patients with DCIS were retrospectively reviewed Statistical comparisons were performed using the chi-square test Results 1 Calcified DCIS showed positive ultrasound US findings in 80% 44 55 of cases The most common US finding was nonmass lesions 43 6% 24 55 Nine cases had pure ductal dilatations 16 4% 9 55 Non-calcified DCIS showed positive US findings in 96 4% 27 28 of cases The most common US finding was mass 89 2% 25 28 Two cases had pure ductal dilatations 7 1 % 2 28 No significant difference was found in the shape margin orientation posterior feature of a mass between the calcified and non-calcified groups P >0 05 Significant difference was observed in the size boundary echogenicity on ultrasound of the two groups P <0 05 2 At histopathology the pathological scores high nuclear grade positive ER status positive PR status positive Ki67 status and the presence of Her-2 neu oncogene were more common in the calcified group than in the non-calcified group Conclusions Calcified and non-calcified pure DCIS have different pathologic and sonographic features Calcified DCIS has more aggressive histological features than non-calcified DCIS.

Objective To study the synergism effect of 131 I-Herceptin and high-energy X-ray on HER2 overexpressed breast cancer SK-BR-3 cells.Methods The protein expression and gene amplification of human epidermal growth factor receptor-2 ( HER2 ) in SK-BR-3 cells were identified by immunohistochemistry and fluorescence in situ hybridization ( FISH ) method, 131 I-Herceptin was prepared by iodogen method, and the IC15 concentration of 131 I-Herceptin on SK-BR-3 cell were selected by MTT method.The cells were divided into control group and drug group according to 131 I-Herceptin used or not, and were delivered five different doses of external irradiation (0,2,4 and 6Gy), and the synergism effect was detected by colonogenic assay.The cells were divided into blank group, drug group(131I-Herceptin), X-ray group(2 Gy external irradiation) and combination group (131I-Herceptin+2 Gy external irradiation), the apoptosis rate and death rate were detected by AO/EB method and cell cycle were detected by flow cytometry.Results The labling rate, radiochemical purity and specific radioactivity of 131 I-Herceptin were 86.8%, 93.9% and 868.3 μci/mg, respectively.The IC15 of 131 I-Herceptin was 15.625μci/mL.131 I-Herceptin and high-energy X-ray significantly reduced surviving fraction ( SF) ( F=628.888,F=964.97,P<0.05) and there were interactions between them (F=113.046,P<0.05).There were significant differences in apoptosis rate and death rate among blank group, drug group, X-ray group and combination group(F=103.324,F=13.33,all P<0.05),and there were significant differences of pairwise comparison (P<0.05).After irradiation and 131I-Herceptin administration, the cell cycle changed obviously from G1-phase to G2-and S-phase.Conclusion 131 I-Herceptin combined with high-energy X-ray has the synergism effect on HER2 overexpressed breast cancer SK-BR-3 cells.

Objective To reassess the value of CT and clinical criteria as prognostic and severity indicators in acute pancreatitis and to investigate the correlation between them.Methods Of 65 cases of acute pancreatitis,the hospitalization days,fevering days and entire complications (including local and systemic complications)were regarded as clinical endpoints.CT criteria included Balthazar's plain CT scan score,necrosis score,CT severity index(CTSI) and London's PSI score.Clinical criteria included Ranson and APACHE Ⅱscore.Using analysis of variance,t-test and multiple linear regression analysis,the correlation between each criteria and the three clinical endpoints was examined as well as the relation between CT and clinical criteria.The power of each criteria and combination of CT and clinical criteria in predicting entire complications of AP was assessed and compared by using ROC analysis.Results The mean scores of PSI,Ranson and APACHE Ⅱamong three groups classified according to entire complications were significantly different.Except Balthazar's plain CT scan criteria,each criteria's mean scores in group with local complications was signifiantly higher than that in group without and entire complications was significantly more in sever group than that in mild group.Mean hospitalization days and fevering days were significantly longer in sever group than that in mild group with Ranson Score.PSI and Ranson score had linear correlation with fevering days,and Ranson score had linear correlation with hospitalization days.In CT criteria,a linear correlation was only found between PSI and Ranson score.ROC analysis showed the Az of Ranson score was the largest,and there was no increase in the Az when CT criteria were added to clinical criteria.Conclusion The predictive value of Ranson score and PSI are superior to that of others.Clinical criteria is superior in predicting systemic complications,whereas CT is superior in predicting local complications.There is no improvement in predicting entire complications of AP when CT criteria are added to clinical criteria.The findings of plain CT scan is found to be some laggard compared with that of clinic.CT scan and short follow-up are important in the evaluation of AP.

The development of orthopedic fixing materials is a most basic and active aspect in orthopedic injury treatment.Their invention and application of early manual splints,gypsum bangs,hot-shaped splints,polymeric bangs and so on,greatly promoted orthopedic fixing performance and varied the strategies of orthopedic injury treatment.By repeated experiments and clinical trials,we developed a new orthopedic fixing material using polymeric materials.Convenient to operate,easy to remove,auto-shaped,X ray transmittable,water proof and durable,it not only satisfies the needs of fixing different parts of the body,but also remedies the defects of earlier fixing materials,and therefore has gained an extensive application as the best fixing material for the treatment of clinical and battle-field bone fractures.Further studies on it will surely better its properties,reduce its cost and lead it to a wider application.

Objective To develop a suit of rapidly-fixed evacuation stretcher for safe and convenient transportation of the wounded. Methods By making use of the expansion-shaping property of a bubble-shaped material produced by the chemical reaction of two kinds of Macromolecule materials, and the restrictive effect of body shape matching cotton fabric fixation-bag, a new type of stretcher was developed. Results It was proved by the maneuver and all kinds of traffic accidents that the field stretcher could provide dependable safeguard for the wounded, and the effect was very well. It was suitable to carry out the second or the third treatment and other transportation schemes. Conclusion This new type of field stretcher is light, portable and easily-operated, and besides, it can be used repeatedly.

Objective To summarize and analyze prenatal ultrasound and postnatal autopsy ifndings in fetuses with urorectal septum malformation sequence (URSMS). Methods An analysis of prenatal ultrsound ifndings and postnatal autopsy features was performed on eleven cases of fetuses with URSMS that were identiifed by ultrasonography at Shenzhen Maternity&Child Healthcare Hospital in the period of January 2003 to December 2012. Results Prenatal ultrasonography showed a large abdominal cystic mass concomitant with imperforate anus in eleven fetuses with URSMS. The cyst contained unilocular or bilocular cystic structures in two fetuses, and trilocular cystic structures in nine fetuses. The cyst was demonstrated as clear acoustic transmission in three fetuses and unclear in eight fetuses. Out of them, seven fetuses had kidney abnormalities, six had ascites, and three had enterolithiasis. The associated systemic abnormalities included tethered cord in two fetuses, single umbilical artery in two fetuses, sacrococcygeal dysplasia in one fetus, and myocardial noncompaction in one fetus. 21-trisomy was found in one fetus by chromosome examination. Eleven cases were all identiifed as female fetuses by autopsy ifndings, including a single perineal opening and ambiguous genitalia with clitoral hypertrophy and labial fusion. The internal genital abnormalities included double vagina or longitudinal vaginal septum in nine fetuses, double uterus or uterus bicornis in ten fetuses and vaginal dysplasia in one fetus. Conclusions URSMS is a complex congenital malformation, which includes abnormalities of the urinary system, reproductive system and gastrointestinal track. An abdominal cystic mass visualized by prenatal ultrasonography might be the distinctive lesion in female with URSMS, and have an important diagnostic value. The kidney abnormalities and ambiguous genitalia can contribute to the diagnosis of URSMS.

ObjectiveTo summarize prenatal ultrasound image features of Cantrell′s syndrome (Cantrell′s pentalogy).MethodsFrom January 2007 to December 2013 in Shenzhen Maternity & Child Healthcare Hospital, there were twenty cases diagnosed and conifrmed by postmortem as Cantrell′s syndrome. The prenatal ultrasound image features and outcomes were analyzed.ResultsAmong the twenty cases, eighteen were singleton and two were twins with the other normal fetus. Prenatal ultrasound image: two obvious ultrasound image features of ectopic heart and hight omphalocele (seventeen cases) or gastroschisis (three cases) were appeared in prenatal ultrasound of twenty cases. Fourteen cases were complete ectopic heart (Ectopic heart was located within the omphalocele in one case, ectopic heart was completely exposed in amniotic lfuid in thirteen cases), six cases were incomplete ectopic heart, three cases were ventricular septal defect in which one was associated with pulmonary stenosis, two cases were single atrium and single ventricle in which one was associated with a single arterial trunk, one case was tetralogy of Fallot, and six cases were without signiifcant heart abnormalities. Associated with other abnormalities: seventeen cases were associated with other abnormalities in twenty cases, in which eleven cases were spinal abnormalities, nine cases were umbilical cord abnormalities (short umbilical cord in six cases, single umbilical artery in two cases, short umbilical cord associated with single umbilical artery in one case), ifve cases were neurological abnormalities (exencephalia in four cases, encephalocele in one case), four cases were limb abnormalities, three cases were cheilopalatognathus, one case was giant bladder. Three cases weren′t associated with other abnormalities. Nuchal Translucency: in eight cases, nuchal translucency were measured in early pregnancy. Nuchal translucency of ifve cases were thick, one case was cervical hygroma in one of the two twins. Chromosome examination results: only three cases had chromosome examination in twenty cases, in which one case was trisomy 21, another two were chromosome normal. Fetal outcome and examination results: in twenty cases, one of the two twins can′t survive after birth, and the other one of two twins was reduced in intrauterine, so the specimen could not be obtained. Eighteen cases of singleton underwent labor induction. The results of postmortem appearance (nineteen cases) and autopsies (ifve cases) were all consistent with prenatal ultrasound scan.ConclusionsEctopic heart and hight omphalocele or gastroschisis appearing at the same time was the characteristic of Cantrell′s syndrome in prenatal ultrasound and other abnormalities may be part of the Cantrell′s syndrome spectrum. It is beneift to detect and diagnose Cantrell′s syndrome by prenatal ultrasound in the ifrst trimester.

Objective In this study,we investigated the hair cell apoptosis and the molecular mechanisms in age-related hearing IOSS mouse cochlea.Methods All animals were assessed by ABR and DPOAE analysis in both ears.TUNEL(Terminal dexynucleotidyl transferase(TdT)mediated dUTP Nick End Labeling)and PI were used to identify DNA fiagments and caspase-3 activities in hair cells.Results The results showed that the nmf308 mice had progressive hair cell loss along with age.The cochlear OHCs were reduced 5%～10% at 1 month and 100% at 3 month in the basal region.Substantial amounts of TUNEL-postive OHCs nuclei were observed at age of 1 month,and activated caspase-3 labeling OHCs were most obviously observed at age of 2 months.Conclusion These results suggest that DNA single strand break is attributed primarily to apoptosis of cochlear lesion,and activation of caspase-3 at leter stage leads to the hair cell apoptiovs of nuclear condensation in age-related hearing loss mouse cochlea.

Three pSIREN-siRNA plasmids were constructed using a pSIREN-RetroQ vector to silence the expression of muhidrug resistance-associated protein (MRP1) gene, and subsequently characterized by restriction endonuclease digestion and DNA sequencing. A truncated MRP1 and a full-length MRP1 were cloned into pEGFP-N2 and PeDNA3.1 respectively as pEGFP-MRP1T and pcDNA-MRP1. The plasmid pEGFP-MRP1T was co-transferred with each of the three pSIREN-siRNAs into HEK293 cells for MRP1T-GFP targeted silencing, and pSIREN-siRNA1 was used as the negative control, pSIREN-siRNA2 and pSIREN-siRNA3 appeared to be more effective to silence MRP1T-GFP compared to pSIREN-siRNA1 as shown by fluorescence microscopy. For the silencing of full-length MRP1 expression, HEK293 ceils were co-transferred with pcDNA-MRP1 and either of the three pSIREN-siRNAs, then subjected for Western blot analysis and MTT assays, pSIREN-siRNA2 and pSIREN-siRNA3 were able to inhibit the expression of 190 kD MRP1, but not pSIREN-siRNA1. The MDR of MRP1-transfected HEK293 ceils was abolished with pSIREN-siRNA2 or pSIREN-siRNA3 transfections. RNA secondary structure predictions demonstrated that the mRNA local free energy (△G) of the siRNA1 targeted sequence was lower, as the GC content and Tm value of siRNA1 were higher than those of siRNA2 and siRNA3. These data suggest that the local structure siRNAs and target mRNA may influence the silencing efficiency of MRP1 expression.