The embryo development technique in Zebrafish, Brachydanio rerio, is a toxicity testing method making use of the high sensitivity of fish embryo development in early stage to study and evaluate the specific effecting mechanism, the most sensitive effecting time, embryo toxicity and teratogenicity of chemicals through observing the development process of zebrafish embryo after chemical exposure to fertilized ova. This technique has been widely used to test toxicity of chemicals with the advantages of low cost, high sensitivity, simple to operate and simultaneous to detect multi-endpoints. The main methodology, technical characteristics and the status of world-wide application of this technique are reviewed in this paper. Based on the urgent environmental problems in China, the prospects to use this method for monitoring toxicity of mixed pollutants in wastewater are put forward.

The study aimed at investigation the effect of the transgenic biocontrol bacteria 308R(pCPP430) on the tomato rhizosphere bacterial communities. Strain 308R (Pantoea agglomerans), an adnascent bacterium isolated from tomato, was a biocontrol bacterium. The recombinant cosmid pCPP430 contained the hrp gene cluster of Erwinia amylovora was transformed into 308R Strain. Transgenic biocontrol bacterium 308R(pCPP430) produced Harpin protein which induce hypersensitive response and disease resistance in plants and, surprisingly, increase plant growth. Two complementary methods, sole-carbon-source utilization tests (SCSU) and ERIC-PCR, were used to compare the characterization of bacterial communities from three groups of tomato roots dipped in sterilized water, 308R suspended liquid and 308R(pCPP430) suspended liquid. The cluster analysis and principal components analysis of the number of colonies from SCSU indicated consistent differences in the rhizosphere of each group. Cluster analysis of SCSU data showed that there were consistent differences in the rhizosphere of each group. The rhizosphere bacteria communities of root dipped with 308R(pCPP 430) and 308R have a relative good replication, but the differences of corresponding replicates in dipped with distilled water was increased by degrees. Cluster analysis of ERIC-PCR fingerprints revealed that most of results (eight in ten) have overlaps between roots dipped in water and those dipped in 308R liquid. The methods used in this study may prove a useful approach for the comparison of bacterial communities.

Purpose:To study the expression of CD44v6 protein in breast carcinoma and its prognostic significance. Methods:100 cases of formalin-fixed paraffin-embeded female breast invasive ductal carcinoma tissues were retrospectively analyzed using EnVision~(TM) immunohistochemical method with the monoclonal antibody CD44v6.Statistical analysis was based on the Log-rank test and Cox analysis.Results:Sixty-six of 100 cancer tissues expressed CD44v6.Positive staining was mainly on the cell membranes.There was significant correlation between CD44v6 immunoreactivity and lymph node me- tastasis and TNM stage.The 5-and 10-year survival rates were 82.76% and 78.37% of patients with CD44v6 low-expres- sion tumors,and 64.1% and 49.88% of those with CD44v6 high-expression tumors,respectively;the difference between the two groups of patients was significant(P

0.05).There were significant differences between groups A and C(P

Objective: To investigate the characteristics of breakthrough cases of varicella in Haishu District, Ningbo City, Zhejiang Province from 2017 to 2022, so as to provide the evidence for varicella prevention and control. Methods: Information on reported cases of varicella and vaccination in Haishu District from 2017 to 2022 were collected through the China Infectious Disease Reporting Management Information System and the Immunization Program Information Management System of Zhejiang Province. The breakthrough cases who had received 1 or 2 doses of varicella vaccine 42 days before the onset of disease were selected. The distribution of sex, age, region and the interval between onset and last immunization were descriptively analyzed. Results: A total of 1 563 varicella cases were reported from 2017 to 2022. There were 928 breakthrough cases (59.37%), of which 660 cases with 1-dose immunization history (42.23%) and 268 cases with 2-dose immunization history (17.15%). The proportion of 1-dose breakthrough cases in total annual cases showed a decreasing trend (P<0.05) from 2017 to 2022, while there was no significant tendency on the proportion of cases with 2-dose immunization history (P>0.05). There were 392 males (59.39%) and 268 females (40.61%) with 1-dose immunization history. There were 150 males (55.97%) and 118 females (44.03%) with 2-dose immunization history. Breakthrough cases with 1- and 2- dose immunization history had an median age of 13.00 (interquartile range, 4.00) and 9.00 (4.00) years, respectively, and the difference was statistically significant (P<0.05). The regional distribution of the breakthrough cases with 1- and 2- dose immunization history was mainly in the rural-urban fringe, with 319 cases (48.33%) and 137 cases (51.12%), respectively, and the difference was statistically significant (P<0.05). The median interval between onset and last immunization was 12.00 (interquartile range, 3.00) and 4.00 (3.00) years, respectively, and the difference was statistically significant (P<0.05). Conclusions The breakthrough cases of varicella in Haishu District from 2017 to 2022 were mainly cases with 1-dose immunization history, males, and residents in rural-urban fringe. The age was older and the interval between onset and the last immunization was longer in cases with 1-dose immunization history than in cases with 2-dose immunization history.

Objective To evaluate the value of captopril challenge test (CCT) in the diagnosis of primary aldosteronism (PA).Methods A total of 674 patients [(45.0±13.7) years, men 341, women 333] admitted to Peking Union Medical College Hospital from 2000 to 2015 were analyzed.Among them, 222 subjects were with essential hypertension (EH), 28 were with pheochromocytoma (PHEO), 246 were with idiopathic hyperaldosteronism (IHA) and 178 were with aldosterone producing adenoma (APA).All patients received CCT.24 h urine sodium was measured in partial patients.Plasma renin activity (PRA), aldosterone (ALD) were detected.Results Compared with EH [PRA: before 0.5(0.2,0.9) μg·L-1·h-1, after 0.8(0.4,1.5) μg·L-1·h-1;ALD: before (393±122) pmol/L, after (360±97) pmol/L] and PHEO [PRA: before 0.3(0.1,0.9) μg·L-1·h-1, after 0.4(0.1,1.6) μg·L-1·h-1;ALD: before (396±108) pmol/L, after (374±114) pmol/L], lower levels of PRA and higher levels of ALD before and after CCT were observed in PA patients [PRA: before 0.1 (0.1,0.2) μg·L-1·h-1, after 0.1 (0.1,0.2) μg·L-1·h-1;ALD: before (468±216) pmol/L;after (457±199) pmol/L].After CCT, the suppression rate of ALD [2.8% (-8.8%,15.4%) vs 6.6% (-4.3%, 17.6%)] and increasing rate of PRA [0(0,50%) vs 50%(0, 200%)] in PA patients were lower than those in EH patients.The ALD/PRA ratio (ARR) were higher in PA than that in EH or PHEO patients.In the EH subjects, ALD levels of seated posture were higher than those of recumbent posture both before and after receiving captopril, but with no changes in ARR after CCT.No significant differences in ALD and ARR (before and after receiving captopril) were observed between seated and recumbent position in the PA group.The ARR after CCT tended to decrease in EH subjects with elevated urine-sodium compared with those with normal urine-sodium.No changes could be viewed in ALD and PRA levels between normal urine-sodium and elevated urine-sodium groups among APA, IHA and EH patients either before or after CCT.Among patients with APA, the ALD levels before CCT and the ARR after CCT were lower in the patients with AngiotensionⅡ(AngⅡ) reactive than those without.A ROC curve analysis suggested that the optimal cutoff value was 46.2 (ALD unit:ng/dl;PRA unit:μg·L-1·h-1) for ARR after challenge in diagnosing PA, with the sensitivity of 88.7% and specificity of 84.8%.Conclusions ARR after 25 mg captopril had high sensitivity and specificity in diagnosis of PA with the cutoff of 46.2.Seated CCT could replace recumbent CCT as a more confirmatory test.The PRA increasing rate should be taken into consideration when diagnosis of PA.

Objective To observe the changes in cardiopulmonary exercise test (CPET) variables of children with frequent premature ventricular contraction (PVC)before and after creatine phosphate treatment,to examine the difference of CPET results between PVC patients and healthy children,and to evaluate the effect of frequent PVC on the cardiac reserve function in children.Methods One hundred and nine frequent PVC children and 98 healthy children underwent treadmill exercise test and CPET respectively,the changes of CPET variables were observed among patients before and after treatment as well as among the healthy children.CPET variables include maximal oxygen consumption/kg (VO2max/kg),maximal oxygen consumption/heart rate( VO2max/HR),HR( at different time point),anaerobic threshold ( AT),and AT/VO2 maximum prediction (AT/VO2max pred).Results Before creatine phosphate treatment,VO2max/kg,VO2max/HR and AT/VO2max in frequent PVC children were ( 22.9±7.4 ) ml/( kg·min),( 9.3 ± 1.5 ) ml,( 15.5 ±2.7 ) ml/( kg· min).After creatine phosphate treatment,VO2 max/kg,VO2 max/HR and AT/VO2 max were (26.4 ± 6.0) ml/( kg· min),( 11.4 ± 3.3 ) ml,and ( 17.4 ± 3.8 ) ml/( kg· min).These CPET variables after creatine phosphate treatment in frequent PVC children were obviously higher than those before treatment and there was a significant difference [ VO2 max/kg ( t =2.11,P ＜ 0.001 ),VO2 max/HR ( t =4.02,P ＜ 0.001 ),AT( t =10.2,P ＜ 0.001 )].Control group of 98 healthy children had negative CPET results.Conclusion Cardiac reserve function decreases in frequent PVC children.After treatment,the exercise capacity and cardiac reserve function can be improved.CPET can reflect cardiac functional reserve changes in PVC children.

ObjectiveTo detect mRNA and protein expression of steroidogenic factor-1 ( SF-1 ) and DAX-1 in human adrenocortical tumors and normal adrenal cortex,and to investigate the effect of SF-1 and DAX-1 on the steroidogenesis and development of adrenocortical tumors.Methods Total RNA and protein was extracted from angiotensin Ⅱ unresponsive aldoterone-producing adenomas ( A Ⅱ -U-APA,n =12 ),angiotensin Ⅱ responsive aldoterone-producing adenomas ( AⅡ -R-APA,n =5 ),cortisol-producing adenomas ( CPA,n =10 ),adrenal nonfunctional adenomas ( NFA,n =10 ),aldosterone-producing carcinoma ( APC,n =2 ) and normal adrenal cortex ( NAC,n =8).To analyze gene expression of SF-1,DAX-1,ACTH receptor(ACTHR),and β-actin by real-time quantitative PCR in different tissues.The protein expression of SF-1,DAX-1,and β-actin in the same tissues by Western blot.To study the relationship of ACTHR,SF-1,and DAX-1 with clinical data in adrenocortical tumors.ResultsThe expression of SF-1,DAX-1 mRNA and protein was different in NAC,AⅡ -U-APA,A Ⅱ -R- APA,APC,CPA,and NFA tissues [ relative expression of SF-1 mRNA:24.58±2.45,23.89±3.17,21.59±3.00,(38.75,44.16),14.17±2.80,and 36.38±3.50; DAX-1 mRNA:0.57±0.06,0.37±0.05,0.43±0.05,( 1.52,1.21 ),0.39 ±0.04,and 0.83 ±0.08 ; SF-1 protein:0.76 ±0.11,0.76 ±0.10,0.73 ±0.07,(1.24,1.40),0.55±0.04,and0.98±0.10; DAX-1 protein:0.65±0.14,0.39±0.13,0.43±0.14,(1.18,1.02),0.56±0.04,and 1.03±0.13 ; all P＜0.05 or P＜0.01 ].There was negative correlation by higher SF-1/DAX-1 ratio and tumor size in AⅡ -U-APA tissues.The mRNA and protein expression of SF-1 was lower in CPA and there was the positive correlation with tumor size.Conclusion SF-1 and DAX-1 might play a key role in the development of the adrenocortical tumorigenesis and steroidogenic tissues.

ObjectiveToestablisharatmodelofincreasedbloodflow-inducedpulmonaryarterialhypertension generatedbyanastomosisoftheleftcommoncarotidarterytoleftexternaljugularvein.Methods 45maleSDratswere divided into three groups:the shunt group , the ligation group and the sham group .At twelve weeks after the procedure , the general status of the rats was observed . Heart conditions , cardiac output and shunt patency were measured by echocardiography .Right ventricular systolic pressure ( RVSP ) and Qp/Qs were checked by catheterization . Right ventricular hypertrophy index ( RVHI) was calculated and lung tissues were examined by pathology using hematoxylin -eosin and elastin Van Gieson staining .All data were analyzed statistically by one-way ANOVA test using SPSS 16.0.Results There was no significant difference in body weight gains between the groups .The patency rate of shunt was 84.6%.The heart was enlarged in the group shunt .Cardiac output increased significantly in the shunt group than that in the other two groups [(309.8 ±33.1) mL/min?kg vs.(245.6 ±31.9) mL/min?kg, (240.8 ±30.9)mL/min?kg, respectively, P<0.05].In the shunt group Qp/Qs was 2.16 ±0.38 and RVSP increased to (35.8 ±4.9) mmHg, RVHI was 0.3263 ± 0.0342, significantly higher than that of the other groups .The pulmonary arteriolar wall was evidently thickened in contrast to that in the sham group [ ( 22.3 ±1.7 )% vs.( 10.6 ±1.7 )%, P <0.05 ) .Conclusions Anastomosis of the left common carotid artery to left external jugular vein can successfully establish pulmonary arterial hypertension model induced by high blood flow in rats .

Objective To investigate the correlation between premature beats,atrioventricular block (AVB),heart rate variability (HRV) and age as well as gender among normal children.Methods One thousand five hundred and eighty-one cases of normal children from Beijing were divided into three age groups:3 to 6-year old,～ 10-year old and ～ 12-year old.All children were evaluated with both ECG and Holter to determine arrhythmia and HRV.The arrhythmia incidence and five time-domain values of HRV were analyzed according to age,gender and the existence of ventricular premature contractions.Results (1) Both ECG and 24-hour ECG monitoring (Holter) data showed that the average heart rate (HR),the slowest HR and maximum HR of normal children decreased with age,while PR interval and QTc interval increased with age.The correlation between age and the four indices were statistically significant (P ＜0.01),except the PR interval.(2) The positive rate of arrhythmia with Holter and ECG in normal children had statistically significant difference(x2 =4.810,P ＜ 0.05).(3) The incidence of premature contraction and Ⅰ ° AVB rised or fell with increasing age(P ＜ 0.05).Compared with 3 ～6-year age group,10-year and ～ 12-year old group had highly risk tendency in premature contraction(OR =1.31,2.04).(4) The five time-domain indices of HRV were increases with age.Each age group had statistically significant difference (P ＜ 0.01).(5) The five time-domain indexes of HRV in males were higher than those in females (P ＜0.01).(6) No significant differences were found with five time-domain indexes in children with or without ventricular premature contraction (P ＞ 0.05).Conclusions (1) HR of normal children decreases with age,but QTc interval increases with age.(2) The supraventricular premature contraction is the most common arrhythmia in normal children with Holter.(3) The occurrence of premature contractions increases with age,more obvious during the preadolescence.Ⅰ ° AVB occurs with age growth on a downward trend.Sex has no effect on the same arrhythmia.(4) HRV in normal children has age difference,suggesting that the pediatric cardiac autonomic nervous system (ANS) was immature.(5) The differences of HRV in sex were most significant during the preadolescence,which showed the developmental inconsistencies in cardiac ANS between male and female of preadolescence.(6) In the majority of normal children,the ventricular premature contractions were benign.