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.

AIM:To establish a quantitative method for determination of hesperidin in Citri reticulatae in order to evaluate their quality. METHODS:Sample was extracted with methanol by means of sonication for 30min. ODS column was used with methanol-water-acetic acid(35∶61∶2)as mobile phase. Detection wavelength was at 284nm. RESULTS:Hesperidin in sample solution was well separated. Linearity was good(r=0.9995)in range of 0.202～2.424?g. The average recovery was 98%, RSD of repeatability was 2.3%. CONCLUSION:This method is simple, accurate and reliable, which may well be used for the determination of hesperidin.

Objective:To investigate the ethical practices in individual supervision of counseling in China.Method:Forty-one graduate students and 38 participants of the China American Psychoanalytic Alliance (CAPA) were included in this study.The Ethical Practices in Supervision Scale (EPSS) was used for the investigation.The scale included two parts.The first part was to investigate supervisors'ethical practices,and the second part focused on supervisees'impact from unethical supervisors.Results:The findings showed that 7 out of 28 items in the scale were significantly different between two groups.CAPA participants' supervisors performed better than graduate students' supervisors (P ＜ 0.05) in supervision scheduling,ethics discussion,and counselor/therapist role.Graduate students' supervisors performed better than CAPA participants' supervisors (P ＜ 0.05) in live supervision,audio/video review,informing clients of supervisee's status as trainee,and supervision contract.If individuals experience their supervisors being unethical or unprofessional,more than 55 percent of the individuals responded to negatively affect their thoughts toward their supervisor and the supervisory alliance,and almost 50 percent of the individuals responded to negatively impact their emotion and quality of their counseling for clients.Conclusion:Both graduate students and CAPA participants respond that their supervisors perform well in both monitoring supervisees' performance and maintaining appropriate boundaries.However,both of them should work more on reviewing video or audiotapes of supervisees' sessions regularly.

Objective A meta-analysis was conducted to evaluate the correlation between hepatitis C virus (HCV) infection and the incidence of intrahepatic cholangiocarcinoma (ICC).Methods EMBASE,MEDLINE,Web of Science,CNKI,Weipu and Wanfang databases were retrieved to identify eligible studies which were published between January 2000 and May 2015.Pooled odds ratios (OR) with 95％ confidence interval (95％ CI) were calculated using RevMAN 5.3.Results 14 case-control studies and 2 cohort studies were included in this study.As there was great heterogeneity among these 16 studies (Chi2 =53.18,df =15,I2 =72％,P ＜0.05),the random-effect model was employed.The combined risk estimates of all the studies showed a significant increase in ICC incidence with HCV infection (OR =3.96,95％ CI 2.63-5.95,P ＜ 0.05).The Begg funnel plot showed no evidence of publication bias.Conclusion HCV infection is related to an increased risk of ICC incidence.

Objective To evaluate the effects of morphine preconditioning on the expression of microRNAs (miRNAs) during hypoxia-reoxygenation (H/R) in isolated cardiomyocytes in rats with heart failure.Methods Healthy adult male Sprague Dawley rats,w eighing 200-220 g,were used in this study.Adriamycin 2.0 mg/kg was injected once a week for 6 weeks via the tail vein to induce heart failure.The cardiomyocytes were isolated from the failing hearts of rats and seeded in 24-well plates or in 60 mm diameter dishes.The cells were then randomly divided into 3 groups (n =16 each) using a random number table:control group (group C); group H/R;morphine preconditioning group (group MP).The cells were cultured in normal culture atmosphere in group C.After being exposed to hypoxic air (5％ CO2-95％ N2) for 90 min,the cells were returned to the high-glucose DMEM supplemented with 10％ newborn bovine serum and were then cultured for 120 min in H/R and MP groups.In group M,the cells were cultured in morphine culture medium (final concentration of morphine 0.3 μmol/L) for 10 min and then were returned to the culture medium without morphine and cultured for 30 min immediately before hypoxia.At 120 min of reoxygenation,the cells of 8 wells in each group were chosen to detect the cell viability and lactate dehydrogenase (LDH) activity (by Typan blue staining).All the RNAs were extracted from the cardiomyocytes of the left 8 wells in each group and subjected to miRNA microarray to screen differentially expressed miRNAs.Results The cell viability was significantly lower,the activity of LDH was higher,the expression of miR-6216 and let7e-5p was higher,and the expression of miR-133b-5p was lower in H/R and MP groups than in group C (P ＜ 0.05).Compared with H/R group,the cell viability was significantly increased,the activity of LDH was decreased,the expression of miR-133b-5p was up-regulated,and the expression of miR-6216 and let-7e-5p was down-regulated in MP group (P ＜ 0.05).Conclusion Morphine preconditioning reduces H/R injury to isolated cardiomyocytes in rats with heart failure through regulating the expression of miRNAs such as miR133b-5p,miR-6216 and let-7e-5p.

BACKGROUND:It is reported that bone marrow mesenchymal stem cell(BMSC)transplantation might be a promising treatment for liver fibrosis.But the mechanism is still unclear.OBJECTIVE:To observe the hepatic stellate cells apoptosis induced by BMSC transplantation,and to study the mechanism of BMSC in treating hepatic fibrosis in vivo.METHODS:CCl_4 subcutaneous injection was performed to induce rat liver fibrosis.After 8 weeks of CCU injection,20 rats which underwent successful model establishment were randomly divided into experimental group and control group,10 in each group.The experimental group received MSC transplantation via tail vein injection,and the control group were given DMEM instead.The rats were killed and the livers were harvested at three time point,the day of MSC transplantation,3 days after transplantation,and 7 days after transplantation.The hydroxyproline content was detected by HE and Masson staining,and the expression changes of α-smooth muscle actin(α-SMA)proteins were determined using immunohistochemistry.The apoptosis of hepatic stellate cells were determined by α-SMA and TUNEL(terminal dUTP nick-end labeling)dual-staining.RESULTS AND CONCLUSION:After 8 weeks of CCU injection,the hydroxyproline content increased and histology indicated progress of liver fibrosis.At 7 days after MSC transplantation,the hydroxyproline in the liver was decreased,and the liver fibrosis was alleviated in the experimental group but aggravated in the control group.Immunohistochemistry indicated that α-SMA positive cells were increased at 8 weeks after CCU injection.At day 7 after transplantation,α-SMA positive cells in the experimental group were significantly less than control group(P ＜ 0.05).At 3 days after transplantation,the hepatic stellate cells apoptosis in the experimental group was significantly aggravated compared with control group(P ＜ 0.05).This suggested that MSC transplant was an effective treatment for liver fibrosis.MSC inducing hepatic stellate cells apoptosis may be one of the mechanisms.

Objective To investigate a poisoning caused by wild mushrooms and to identify the toxin in these mushrooms.Methods Epidemiological investigation,blood test and mushroom toxin were analyzed.Results This incident was taken place in one family,and all family members were dead.Multiple organ damage was observed in all patients;amatoxins and virotoxins were detected in both mushrooms and the soup,but were not detected in blood samples because of dialysis.Conclusion The incident was caused by wild mushrooms and public education shoud be strenthened to urge people to avoid eating wild mushrooms and go to the hospital immediately if poisoning takes place.

Objective To explore a new modified stable genioplasty with modified MortiseTenon joints technique and to analyze the surgicaI results in clinic as well as to discuss the advantages in theory.Methods Eleven patients were performed by the genioplasty with Mortise-Tenon joints technique intraoral approach from June 2012 to March 2014,who were suffered from retrusion of chin with different degree accompanying with long chin.The two segments of the chin by osteotomy were made into the Mortise-Tenon joints and fixed by titanium plates.The results were evaluated by clinical appearances and image analyses.Results All 11 patients were satisfied with the procedure and the contour of the chin.The surgical effects were stable without any complications.The advancement in average Mortise-Tenon joints technique was 7.38 mm.Conclusions Much better in stablility and less relapse are achieved by using modified Mortise-Tenon joints technique;furthermore,the genioglossal muscle and geniohyoid muscle are reserved as much as possible,and therefore the normal function of them could be maintained.This method is a good alternative technique for chin retrusion coexisted withlong chin.

Objective:To explore the technique of two-flap in ear reconstruction.Method:Quantitative tissue expansions were used in the mastoid area in the first stage.After the final injection,there was 1 month of sustaining time.Expanded skin flap and unexpanded fascia flap were designed in the second stage,so thetwo-flaptechnique was used in the ear reconstruction.From January 2004 to December 2008,1 427 patients of microtia were treated using two-flap technque.Result:The expanded skin flap could show the fine structures of the reconstructed ears.The reconstructed ears had vivid cranioauricular angle after using the unexpanded fascia flap.Conclusion: Two-flap method was easily manipulated and the complications were rare.The reconstructed ears had lucid and three-dimensional contour.

AIM:To explore the changes of plasma levels of soluble vascular endothelial growth factor receptor 2 ( sVEGFR2) and superoxide dismutase ( SOD) in hypertensive patients and hypertensive diabetic patients.METHODS:In this cross-sectional study, 88 cases were enrolled, which were divided into hypertensive group (n=31), hypertensive diabetic group ( n=31 ) and control group ( n=26 ) .Blood pressure was obtained from each participant with mercury sphygmomanometer.The levels of sVEGFR2 and SOD were measured by ELISA.Meanwhile, the levels of plasma glucose, glycosylated hemoglobin A1c ( GHbA1c) and lipid profile were detected.RESULTS:The levels of total cholesterol ( TC) and body mass index (BMI) were significantly higher in hypertensive group than those in control group (P<0.05).The levels of TC, low-density lipoprotein cholesterol ( LDL-C) , triglyceride ( TG) , BMI, waist circumference were significantly higher in hypertensive diabetic group than those in control group (P<0.05).The plasma levels of sVEGFR2 and SOD in both hypertensive diabetic group and hypertensive group were significantly decreased compared with control group ( P<0.05), while the mean plasma levels of sVEGFR2 and SOD in hypertensive diabetic group were significantly decreased compared to the hypertensive group ( P<0.05 ) .A significantly positive correlation between sVEGFR2 and SOD in the whole study population (P<0.05) was observed.CONCLUSION: The plasma level of sVEGFR2 is decreased in both hypertensive and hypertensive diabetic patients, and more significantly decreased in hypertensive diabetic patients.De-creased SOD level may be associated with to the reduction of sVEGFR2.