Objective To discuss the application o immunohistochemistry autostainer in immunohistochemistry.Methods More than 30 kinds of first antibodies of the Pathology Department,such as AAT,CerBb-2,CK,HBcAg,ER,PR,Ki-67,and unified second antibodies were selected.Immunohistochemistry autostainer and manual operation were applied to the staining of the antibodies,and then the results by the above methods were compared.Results The antibodies stained by immunohistochemistry autostainer,gifted with clear background,no edge effect,uniform staining and accurate positive results,were all better than those by manual operation from all aspects.Conclusion Immunohistochemistry autostainer is highly automatic,time-saving,manpower-saving,repeatable and highly standardized.

Obstructive jaundice is a common disease that greatly threatens human health.In order to better investigate the pathogenesis of obstructive jaundice,pathophysiological changes during disease progression,and treatment measures,the establishment of a stable animal model of obstructive jaundice becomes the basis for the research on various types of obstructive jaundice.This article elaborates on the advantages and disadvantages of the animal models of acute,progressive,chronic fluctuant,malignant,and reducible obstructive jaundice,as well as their application in clinical practice.These animal models have certain limitations in reflecting the development and progression of target disease.Most animal models are established by surgery or external physical and chemical damage,and there are still no mouse models of gene knockout or overexpression.Future research will focus on the stable animal models of chronic and progressive obstructive jaundice and special types of obstructive jaundice.

Objective To explore the application of iPad-based software Let'S Talk in the rehabilitation of children with autism. Meth-ods From March to September, 2015, iPad-based software Let'S Talk was applied in four children with autism. The language ability, emo-tion and behavior control ability, self-consciousness and eye contact were observed. The Psycho-educational Profile-3rd (PEP-3), Autism Be-havior Checklist (ABC) and Childhood Autism Rating Scale (CARS) were used to access the months of age, behavior characteristics, and the severity of autism before and after application. Results The language ability and behavior control ability improved, as well as the self-consciousness and eye contact, and the bad behaviors reduced. The months of age rose in PEP-3, and the scores of ABC and CARS de-creased. Conclusion IPad-based software Let'S Talk could be applied in the rehabilitation of children with autism.

Objective To assess the value of ultrasonography in distinguishing pharyngoesophageal diverticulum from thyroid nodule.Methods High-frequency sonography was used to detect the size,shape, echo and blood flow of cervix masses in 1219 patients in a lateral decubitus position after drinking water. Results On enhanced power,the image changing rates of pharyngcesophageal diverticulum and thyroid nodule were 71.43% and 14.19% respectively,and their difference was statistically significant(P<0.05). On drinking water,the image change rates of pharyngoesophageal diverticulum and thymid nodule were 100.00% and 1.98% respectively,and their difference was statisfically significant(P<0.05).The detection rates for pharyngoesophageal diverticulum and thyroid nodule were 0.098% and 17.042% respectively. Conclusions Ultrasound examination is of value in distinguishing pharyngoesophageal diverticulum from thyroid nodule in general health check up and regular health examination.

Objective To investigate the arousal effect in the emotional Stroop effect by systematically manipulating the valence and arousal of affective stimuli.Methods 27 college students were recruited to distinThe main effect of emotion on RT was significant (F(5.130) = 5.90, P ＜ 0.01 ), RTs in positive (( 893±36 ) ms)main effect of Emotion on ACC was marginally significant (F(5.130) = 2.63, P = 0.05 ), ACC in high arousing negalence on RT was significant (F(1.26 = 7.03, P =0.013 ).Further analysis revealed that RTs in positive condition was significant (F(1.26) =5.63, P=0.025),ACC in high arousing condition (0.93 ±0.17) was lower than ACC in low arousing condition (0.95 ± 0.09 ).Conclusion The emotional Stroop effect mainly depends on the arousal information of affective stimuli.

Objective To explore the safety and feasibility of controlled hypotension with urapidil and nitroglycerin esmolol-three drugs combination in endoscopic surgery. Methods Forty patients who were received endoscopic surgeries were randomly divided into two groups, 20 cases in each group, urapidil-esmolol-nitroglycerin controlled hypotension group (A group) and non controlled hypotension group (B group). Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), and heart rate (HR) were recorded at the following points: before anesthesia (basal, T1), immediate before surgery (T2), 30 minutes after the start of surgery (T3), end of the operation (T4), 10 minutes after the end of surgery (T5). We recorded blood loss, operative time, transfusion volume, urine. At last the parameters were compared between the two groups. Results The difference of SBP, DBP, MAP at the points of T2, T3, T4 in the two groups was statistically significant (P < 0.05) comparing with the control group (group B). The vital signs of patients during anesthesia in A group had more stable than that of the B group , and the operation time of the A group was significantly shorter than that of the B group. Meanwhile, the blood loss in A group was significantly decreased(P < 0.05). Conclusion It has synergistic antihypertensive effect, and can reduce both side effects and the risk of anesthesia and surgery for us to use urapidil-esmolol-nitroglycerin three antihypertensive drugs in combination. It is a safe and reliable method for clinical application.

Objective To establish chromatographic fingerprint of water and ethanol soluble component of Radix Dipsaci by using HPLC for identifying and evaluating its quality. Methods HPLC method was set up with Agilent HC-C18 column. The mobile phase was comprised of acetonitrile(A) and 0.05mol/L KH2PO4-H3PO4 buffer solution pH=3.0(B) in gradient elution mode. UV detection wavelength was at 220 nm,with a flow rate of 1.0 mL/min. Results Ten batches of Radix Dipsaci samples from different habitats and harvested in different times were analyzed. HPLC fingerprint of medicinal material of Radix Dipsaci was established preliminarily. Seventeen characteristic peaks were confirmed and the fingerprints of different samples were compared by similarity evaluation software. Conclusion This method shows high precision and good repeatability,so it can be used to assess the quality of Radix Dipsaci.

AIM: To optimize the formulation in the preparation of the Breviscapine Liposomes. METHODS: Using film evaporation-extrusion method to prepare Breviscapine Liposomes according to the uniform design,a optimum formulation was established by determining the entrapment efficiency and the ratio of loading drug. RESULTS: The entrapment efficiency and the ratio of loading drug of Breviscapine Liposomes prepared with cholesterol and lecithin were determined to be 69.60% and 29.06%,respectively.The average diameter is 105.6 nm. CONCLUSION: The application of the uniform design is useful to achieve a large entrapment efficiency and ratio of loading drug.

OBJECTIVE:To optimize the extraction technology of chlorogenic acid in Prunus armeniaca flos,and compare the contents in P. armeniaca flos from different origins and varieties. METHODS:HPLC was conducted to determine the content of chlorogenic acid in P. armeniaca flos;using the volume fraction of ethanol,the ratio of material to liquid,ultrasonic extraction times and time as factor,the content of chlorogenic acid as index,single factor and orthogonal test were designed to optimize the extraction technology,and verification tests were carried out. The optimized extraction technology was used to extract and compare the contents of chlorogenic acid in Armeniaca sibirica from 7 origins of P. armeniaca flos and 3 origins of Armeniaca sibirica flos. RESULTS:The optimized extraction technology was extracting twice with 12-fold 75% ethanol,30 min each time. Under the con-ditions,the content of chlorogenic acid can reach 77.38 mg/g(RSD=0.58%,n=3);the contents of chlorogenic acid in A. sibiri-ca flos and P. armeniaca flos were 77.38-83.33 mg/g and 63.12-70.22 mg/g,respectively. CONCLUSIONS:The established extrac-tion technology is reasonable,stable and feasible. The contents of chlorogenic acid in A. sibirica are higher than that in P. armenia-ca flos;the contents have no obvious differences in the same variety of A. sibirica from different origins.

Background Transforming growth factor-β1(TGF-β1) plays an important role in corneal woundhealing.The effects of TGF-β1 on the synthesis of extra cellular matrix (ECM) vary upon different concentrations.Previous studies focused on the effects of high concentration of TGF-β1 on keratocytes under the two-dimensional culture condition,and the effect of low concentration of TGF-β1 on the synthesis of ECM in keratocytes remains unclear.Objective This study was to investigate the growth of Pellet,a three-dimensional model of corneal stroma cells in vitro,and its ECM synthesis under a low concentration of TGF-β1.Methods Bovine corneal stromal cells were isolated from fresh bovine eyeballs by two-step digestion by collagenase and cultured using DMEM/F12 medium with 10％ fetal bovine serum (FBS).Pellets derived fresh bovine keratocytes with culture medium containing 0.25 ng/ml TGF-β1 +5％ FBS and 0.50 ng/ml TGF-β1 +5％ FBS were established,respectively.The morphology of Pellets was observed under the natural light at 48 hours,1 week,2 weeks and 3 weeks after culture.In 3 weeks after culture,the cell structures was observed by hematoxylin-eosin staining,and Calcein-AM/propidium (Calcein-AM/PI) staining was used to assay the cell viability.Real-time flurorescence quantitative PCR and immunofluorescence technology were applied to analyze the expressions of α-smooth muscle actin (α-SMA),fibronectin (FN),type Ⅰ collagen (Col Ⅰ) and type Ⅲ collagen (Col Ⅲ) mRNA and proteins.RT-PCR was employed to detect the expressions of lumican (LUM) mRNA and keratocan (KERA) mRNA in the cells.Results Cells in Pellet clustered throughout the culture duration.Hematoxylin-eosin staining showed the mass red-dyed collagen fibers in both 0.25 ng/ml TGF-β1 +5％ FBS group and 0.50 ng/ml TGF-β1 +5％ FBS group,and most cells possessed complete structures.The death rate of the cells was (33.60±1.65)％ in the 0.25 ng/ml TGF-β1 +5％ FBS group and (30.90±0.78) ％ in the 0.50 ng/ml TGF-β1 +5％ FBS group,showing an insignificant difference between them (t =0.144,P=0.887).The expressions of α-SMA,FN and Col Ⅲ proteins in 0.25 ng/ml TGF-β1 +5％ FBS group were lower than those in the 0.50 ng/ml TGF-β1 + 5 ％ FBS group (tα-SMA =4.622,P =0.010;tFN =2.973,P =0.040;tCol Ⅲ =7.845,P＜0.001),but the expression of Col Ⅰ in 0.25 ng/ml TGF-β1 +5％ FBS group was higher than that in 0.50 ng/ml TGF-β1+5％ FBS group (tColⅠ =4.022,P=0.016).The ratio of Col Ⅲ/Col Ⅰ in 0.25 ng/ml TGF-β1+5％ FBS group was lower than that in the 0.50 ng/ml TGF-β1 +5％ FBS group in both mRNA and protein level (tmRNA =-3.039,P =0.038;tprotein =3.215,P =0.032).The expression of LUM mRNA and KERA mRNA were detected in Pellet at different time points.The expression of LUM mRNA in 0.25 ng/ml TGF-β1 +5％ FBS group increased over time.While in 0.50 ng/ml TGF-β1 +5％ FBS group,the expression of LUM mRNA peaked at 1 week but declined at 2 weeks.The expression of KERA mRNA in two groups were all peaked at 1 week but declined at 2 weeks.Conclusions Low-dose TGF-β1 in Pellet can maintain the normal growth of keratocytes and synthesize ECM.The expression of ECM tends to the normal condition after reducing the concentration of TGF-β1,implying a scarless expression.