OBJECTIVE To search a method for identifying Clostridium perfringens and genotyping their toxin for gene diagnosis by multiplex PCR.METHODS The mutiplex PCR was developed with three sets of primers(designed) based on the sequences of three C.perfringens toxin genes(CP?,CP? and CPE) published in GenBank to identify C.perfringens and genotype their three toxin genes.RESULTS Three expected(sequences) were (obtained) successfully by multiplex PCR and identified by electrophoresis.CONCLUSIONS The(specific) sequences of C.perfringens could be amplified and their three genes of toxins could be identified by this multiplex PCR(system).Such method should be helpful for developing gene diagnosis well.

OBJECTIVE To discuss a highly effective method to immobilize probe on the surfaces of piezoelectric DNA sensors.METHODS Pseudomonas aeruginosa probe was immobilized on the gold surface of gene sensor(array) with routine self-assembly method(SAM)(non-reduction method) and SAM with deoxidized probe((reduction) method),respectively.The changes in frequency and time-cost were compared in reactions with(different) concentrations of probe.RESULTS Reduction method had the advantage of more probe immobilization;less time consumed in testing and higher changes in frequency during the reaction than non-reduction method.CONCLUSIONS Reduction method has a better ability to immobilize probe on the surfaces of piezoelectric DNA sensors.

OBJECTIVE To discuss the feasibility of signal amplification method with cationic conjugated polymers(liposome) applied during the detection of Pseudomonas aeruginosa using piezoelectric gene biosensors.(METHODS) Oligonucleotide probe for P.aeruginosa was immobilized on the surface of gene sensor array and(hybridized) by PCR production of P.aeruginosa.After hybridization,liposome was added.The frequency shifts were recorded and compared with those ones of the control groups.RESULTS The frequency shifts were(significantly) increasing when adding liposome and the compatible concentration of liposome was 0.8?g/?l.(CONCLUSIONS) Liposome signal amplification is proved to be an effective method to amplify the piezoelectric(signal).

Objective To investigate the cognition and behaviors of sun-protection and the facial photoaging in a Nanjing population, and to analyze the relationships between them. Methods The objects being investigated in Nanjing (n=974) were divided into 10 groups according to age. The ordinary information, knowledge and behavior of sun-protection and Glogau photoageing type of face were studied by questionnaires. The results were analyzed by a logistic regression model to select the related factors to photoageing. Results The risk of skin photoageing increased with age. Most of the 46-65 years old crowds were type Ⅲ photoageing. Most of the objects being investigated had some knowledge and active awareness of sun-protection and could use some ways to protect themselves from sun. However, most people did not use the sunscreen correctly. Those who had higher level awareness and knowledge of sunprotection suffered less risk from skin photoageing. Sunbath without sunscreen for a long time outside activity was a high risk factor of development to skin photoageing. Shade, broad-brimmed hat and sun-protection in autumn were the protective factors. Conclusion Active awareness and correct methods could help prevent skin photoageing. In order to avoid the damages to skin from ultraviolet efficiently, the accurate ways of using sunscreens should be well understood.

OBJECTIVE To establish a SNP detection method by DNA piezoelectric biosensor and detect a SNP relative to HBV infection. METHODS To establish a model experiment with synthesis DNA sequences as target and find the lowest sensitivity. After extraction of genome DNA from inpatient blood sample, the SNP sites located in ESR1 gene region in samples were detected by SNP detecting method established. RESULTS The frequency shift of target-A was 416.0?21.5Hz, the frequency shift of target-G was 9.4?5.0Hz. And it could be detected that the lowest sensitivity of target-A was 2?10-11 mol/L. The three genotypes of blood samples, TT, TC and CC, had different frequency shifts, 109.4?13.4Hz, 52.0?11.4Hz and 7.2?4.5Hz, respectively. CONCLUSIONS SNP in blood sample could be detected specifically and sensitively by DNA piezoelectric biosensor.

Objective To analyze the changes and characteristics of cancer incidence and mortality among Jiaxing residents in the past 6 years,and to provide evidence for the prevention and control of cancer incidence and mortality.Methods Registered malignant tumor data from the Zhejiang Chronic Disease Information Monitoring and Management System were collected and analyzed based on the International Statistical Classification of Diseases (ICD-10).The statistical indexes used for analyses included crude incidence,standardized incidence,crude ortality,standardized mortality,potential years of life lost (PYLL),average years of life lost (AYLL),rate of potential years of life (PYLL‰) and so on.Results From 2010 to 2015,the crude incidence,the standardized incidence,the crude mortality,and the standardized mortality were 342.52/105,240.89/105,194.71/105 and 126.27/105,respectively.The crude incidence and mortality for males were both higher than those for females(378.44/105 vs.307.49/105,x2=763.06,P＜0.01;255.31/105vs.135.62/105,x2-3 814.11,P＜0.01).The PYLL,AYLL and PYLL‰ of residents in Jiaxing were 248,307 years,6.16 years and 12.00‰,respectively.The PYLL‰for males was significantly higher than that for females(x2=37073.85,P＜0.01).The AYLL for females was 6.50 years,higher than that for males(5.98 years)(t =125.15,P＜0.01).The incidence of malignant tumors increased with age,especially in the older than 40 age group,with the older than 80 age group showing the highest rate at 1 779.01/105 (x2 =38 840 907,P ＜ 0.01).Rates for Lung cancer (70.59/105),colorectal cancer (39.86/105),liver cancer (29.14/105),breast cancer(26.73/105) and stomach cancer (26.00/105) were the most common malignant tumors in Jiaxing.Conclusions The morbidity and mortality of malignant tumors are showing signs of rising and the incidences of lung cancer and gastrointestinal cancer are particularly pronounced in Jiaxing.In consequence,measures for the prevention and control of malignant tumors should be developed with clearly defined objectives and research on the pathogenesis of malignant tumors should be stepped upto reduce the incidence and mortality of malignant tumors.

Objective To develop a new type of piezoelectric quartz crystal microbalance DNA sensor based on lambda exonuclease to detect Staphylococcus aureus (S. aureus) and optimize the main detection conditions. Methods After the DNA was extracted from the stain of S. aureus and the target fragment was amplified with self-designed universal primers, the PCR products were treated with Lambda exonuclease and visualized by agarose gel electrophoresis with ethidium bromide followed by a gel documentation system analysis. Then the products with or without exonuclease digestion were added into our piezoelectric quartz crystal microbalance DNA sensor for hybridization. The temperature and time of hybridization were optimized respectively. The specificity and sensitivity of our detection system were evaluated. Results The optimal temperature for hybridization was 35℃ and the optimal time was 60 min. The sensitivity of the lambda exonuclease based DNA sensor was better than that of common DNA sensor. The lowest detection limit of this new type quartz crystal microbalance system was 1.0?104 CFU/ml. Conclusion The lambda exonuclease based quartz crystal microbalance system we designed has the advantages of high efficiency in hybridization, easy to operate, and good response performance. Thus it can be applied to detect S. aureus infections.

Objective To investigate the calculation method and its inlfuencing factors of Z scores in the aortic root diameters measured by echocardiography in children. Methods A total of 105 children with median age 19 months, who came to Shenzhen Children′s Hospital from March 2012 to October 2012 were included. The diameters of aortic ring (ARD) and aortic sinus (ASD) were measured by two dimension echocardiography, Z scores of ARD and ASD were calculated using two different normal reference values regression equation and mean square error derived from Shenzhen children′s hospital (C method) and Pettersen et al (P method). The regression equation from C method and body surface area (BSA) formula from P method were adopted to calculate Z scores of ARD and ASD (ZH method). The Z results of ARD and ASD calculated by those three methods were compared and were analyzed for their normality probability distributions. Results Z scores of ARD and ASD derived from C method were all showed as normal distribution (P=0.067 and 0.650). Z scores of ARD and ASD derived from P method were all showed as normal distribution (P=0.208 and 0.970). Z score of ARD derived from ZH method was showed as non-normal distribution (P=0.027), but Z score of ASD was normal distribution (P=0.430). There were no significant differences in ARD-Z calculated by C method (0.41±0.89), P method (0.23±0.85) and ZH method (0.36±0.94) (F=1.117, P=0.309). There were signiifcant differences in the Z scores of ASD calculated by C method (0.38±0.89), P method (0.58±0.71) and ZH method (0.36±0.84) (F=5.443, P=0.005). Z scores of ARD (r=0.917, P=0.000) and ASD (r=0.900, P=0.000) calculated by C method correlated well with that by P method. Conclusions Calculation method of BSA and normal reference values regression equation were the main influencing factors of Z score value in quantifying children aortic root diameters by echocardiography. For the clinical applications. The normal reference value should be used which is suitable for the Chinese children.

Objective To evaluate the value and clinical application of cardiac structure Z scores in children with preoperative tetralogy of Fallot (TOF) by echocardiography. Methods Considering the result of 31 patients who underwent echocardiography in Shenzhen Children’s Hospital before TOF repair from January 2011 to September 2013, the median age was 10 months (range:3 days to 14 years). Other 31 normal children were selected as normal control group with age-and sex-matched, the median age was 10 months (range: 6 days to 13 years). Echocardiographic measurements included the diameter of right ventricular outflow tract (RVOT), pulmonary valve diameter (PVD), left pulmonary artery diameter (LPAD), right pulmonary artery diameter (RPAD), pulmonary valve peak velocity (PV-Vmax), tricuspid annulus diameter (TVD), mitral annulus diameter (MAD), left ventricular end-diastolic volume (LVEDV) and left ventricular mass (LVmass). Shenzhen Children’s Hospital pediatric echocardiography normal reference values and the regression equation are applied to obtain the forecast average, and using the Z-score formula into the standard value (Z=[M-y]/ MSE ). Results There was no significant difference in body surface area between the TOF group and control group (P>0.05). Compared with control group, the true value of RVOT reduced significantly in TOF group, its Z-score showed negative values increase (-4.87±1.86 vs 0.33±0.93, t=-13.90, P<0.05). PVD, LPAD and RPAD reduced significantly in TOF group, those Z scores showed a negative value increase (-4.15±2.93 vs 0.11±0.85, t=-7.76;-1.35±2.38 vs 0.66±1.17, t=-4.24;-1.47±2.03 vs 0.59±1.04, t=-4.94;all P<0.05). While PV-Vmax increased significantly, and it’s Z score showed a positive value increase (8.38±0.19 vs 0.24±0.98, t=29.50, P<0.05). TVD of TOF patients was slightly larger than the control group (P=0.10), but the Z score was significant greater than the control group (1.00±1.17 vs 0.29±0.52, t=3.06, P<0.05). MV-D, LVEDV and LVmass of TOF patients were slightly decreased than the control group (all P>0.05), but the Z scores of TOF were significantly reduced than the control group (0.25±0.78 vs 0.76±0.65, t=-2.75;-0.92±1.94 vs 0.03±1.01, t=-2.41;-0.83±1.59 vs 0.67±0.69, t=-4.71; all P<0.05). The Z scores of RVOT, PVD, LPAD and RPAD had no significant difference between transannular patch surgery and pulmonary valve-sparing repair patients (-4.89±2.03 vs -4.84±1.67, t=-0.08; -4.73±3.49 vs -3.34±1.75, t=-1.31; -1.88±2.54 vs -0.62±2.00, t=-1.49;-1.89±1.90 vs-0.84±2.15, t=-1.41;all P>0.05). For the Z scores of PV-Vmax, there were no significant difference between transannular patch surgery and pulmonary valve-sparing repair (8.58±1.20 vs 8.12±1.16, t=1.07, P>0.05). Conclusions The Z scores value of cardiac structure can be used in quantitative analysis of RVOT obstruction and development of left ventricular backward of TOF. The Z score of transannular patch surgery is more lower than pulmonary valve-sparing repair patients. The normalized Z score by body surface area correction is important for the preoperative diagnosis and the surgical planning.

Objective To investigate the accuracy of three-dimensional ultrasound volumetric measurements for children's bladder. Methods The bladder volumes of 238 children aged 9 months to 16 years (male:106;female:132) in Shenzhen Children's Hospital underwent real-time three-dimensional ultrasound for volume measurements. These cases were divided into six groups according to the body surface area:(0.36~0.60) m2 (27 cases), (0.61~0.78) m2 (66 cases), (0.79~0.89) m2 (52 cases), (0.90~1.05) m2 (33 cases), (1.06~1.20) m2 (30 cases) and (1.21~1.78) m2 (30 cases). The bladder volume measured by GE Voluson E8 and Philips iU22 were compared with the actual volumes of discharge urine. The relation between bladder volume measurements and the actual volume of discharge urine was analyzed. Results Children's bladder volume increasesd with the body surface area (F=33.53, P<0.05) and showed positive correlations with the body surface area (r=0.679, P <0.05). Compared with the actual volume of discharge urine, each group's bladder volume measurements obtained by three-dimensional ultrasound were lower and the results were as follows:[(47.80±30.33) ml vs (82.24±49.63) ml, (77.20±39.72) ml vs (139.98±79.03) ml, (95.96±50.79) ml vs (175.96±101.70) ml, (117.46±54.17) ml vs (206.62±86.22) ml, (145.53±73.60) ml vs (253.33±135.09) ml and (220.27±110.34) ml vs (327.42±165.45) ml]. The differences between children's bladder volume measurements and the actual volume of discharge urine were statistically significant (t=3.19, 5.53, 5.08, 4.49, 3.84 and 3.00, all P <0.05), but children's bladder volume showed positive correlations with the actual urine volume (r=0.881, P<0.05). The bladder volume measurements by spheroid formula based on two-dimensional ultrasound are lower than the actual volume of discharge urine, and the results were as follows:[(50.38±36.94) ml vs (82.24±49.63) ml, (86.77±62.34) ml vs (139.98±79.03) ml, (102.69±60.21) ml vs (175.96±101.70) ml, (107.94±55.14) ml vs (206.62±86.22) ml, (145.31±66.01) ml vs (253.33±135.09) ml and (222.77±132.59) ml vs (327.42±165.45) ml]. The differences were statistically significant (t=3.03, 4.01, 4.47, 4.95, 3.94 and 2.75, all P <0.05), but children's bladder volume showed positive correlations with actual urine volume (r=0.326, P <0.05). Conclusions Three-dimensional ultrasound volumetric measurements for children's bladder is feasible and in clinical application, the factors of children's growth should be take into account. Combing regression equation, children's bladder volume can be more accurately quantified.