Aim To investigate whether the polypeptide from Chlamys farreri(PCF)protected HaCaT cells from UVB-induced apoptosis through Fas-caspase-3 and ROS-cytochrome C.Methods Experiment designs were divided into five groups:control group,UVB model group,UVB+5.69 mmol?L-1PCF group,UVB+2.84 mmol?L-1 PCF group,UVB+1.42 mmol?L-1 PCF group.SiRNA for Fas inhibited Fas expression of UVB-induced HaCaT cells.Using agarose gel electrophoresis,the effects of Fas siRNA and ROS scavenger NAC on UVB-induced apoptosis of HaCaT cells were investigated.Expression levels of cytochrome C andcaspase-3 after inhibitory Fas were determined by Western blot analysis.Intracellular ROS was detected by means of an oxidation-sensitive fluorescent probe(DCFH-DA).Results SiRNA for Fas had inhibitory effects on UVB-induced apoptosis of HaCaT cells and caspase-3 expression.NAC had inhibitory effects on UVB-induced apoptosis of HaCaT cells.PCF inhibited UVB-induced generation of ROS and cytochrome C release dose-dependently.Conclusions PCF protected HaCaT cells from UVB-induced apoptosis.Its inhibitory effect on apoptosis could be attributed to inhibition of Fas-caspase-3 and ROS-cytochrome C pathways.

Objective To provide a basis for further exploration of Dunaliella Salina by determining the main active components in the extract of Dunaliella salina. Methods The carotenoids and vitamins were determined by HPLC or colorimetric method, amino acids were analyzed by amino acid analyze. Unsaturated fatty acid was determined by GC. Atomic absorption spectrophotometry was used to detect K, Na ,Ca, Mg, Cu, Zn, P, Fe, Mn and SPF was used to detect Se. Results There were affluent carotenoids, vitamins, amino acids, unsaturated fatty acid and trace elements in the extract of Dunaliella salina. Conclusion The extract of Dunaliella salina could be used as natural raw material to develop the supplement food.

Objective To study the effects of polypeptide from Chlamys farreri(PCF) on PI3K/Akt and ASK1-JNK signal transduction pathway in thymocytes after ultraviolet B radiation.Methods Murine thymocytes were exposed to UVB radiation.The reactive oxygen species(ROS) level in thymocytes was detected by colormatching.The activation of Akt was investigated by western-blot.ASK1,JNK activation,mitochondria memberine potential and DNA ladder were also investigated after pretreatment with or without PI3K/Akt pathway inhibitor LY294002.Results PCF could activate Akt,inhibit the activation of the ASK1 apoptosis pathway in murine thymocyte radiated by UVB.Conclusion PCF decreased ROS level,increased Akt activity and inducing ASK1 degradation leading to the inhibition of ASK1-JNK induced apoptosis.

Objective To investigate the effects and mechanisms of all-trans retinoic acid(ATRA)on the proliferation and cell cycle of lung carcinoma cell line A549.Methods The A549 cells were treated with ATRA at the dosages of 5,10,50 ?mol/L for 1-7 d.The proliferation of A549 was assessed by MTT method and cell cycle was analyzed by flow cytometry.The expressions of CDK4,Rb and p-ERK1/2 were assessed by Western blotting.CyclinD1 mRNA was analyzed by SYBR-PCR amplification.Results ATRA obviously inhibited the proliferation of A549 cells,and the cell cycle was arrested in G0/G1 phase.The expression of p-ERK1/2 protein and CyclinD1 mRNA on A549 cells were decreased.Conclusion ATRA might inhibit the proliferation of A549 cells through down-regulating p-ERK1/2 protein and CyclinD1 mRNA.

Objective:To investigate the inhibitory effect and the related mechanism of Yiqi Jianpi Fang on aberrant crypt focus (ACF) in Wister rat colorectal cancer model.Methods:50 Wistar rats were randomly divided into 5 groups:TCM low dose group(TCM solution diluted 3 times,5 mL/kg daily gastric volume),TCM middle dose group (5 mL/kg daily gastric volume),TCM high dose group (15 mL/kg daily gastric volume),model group,normal group.With 10 rats in each group.The colorectal tissues were observed under microscope after methylene blue staining by immunohistochemistry method.Results:Compared with the model group,the number of ACF and large ACF in each TCM group were decreased (P＜0.05),and the number of ACF in the TCM middle dose group reduced most obvious,and the difference was statistically significant (P＜0.05).The proliferation indexs of PCNA in intestinal gland cells 24 h and 48 h after modeling in the TCM groups were higher than those in the model group,and the difference was statistically significant(P＜0.05).The apoptosis index of intestinal gland cells 24 h and 48 h after modeling in the TCM groups were higher than those in the model group,and the difference was statistically significant(P＜0.05).The ectopic expression of β-catenin in TCM groups were lower than that in the model group,and it was highest in the high dose group,than was the low dose group,and the middle group was lowest(P＜0.05).The expression of MMP-7 in TCM groups were lower than that in the model group,and it was highest in the low dose group,than was the high dose group,and the middle dose group was lowest (P＜0.05).Conclusion:Yiqi Jianpi Fang can significantly reduce the number of ACF in Wister rats,inhibit the activation of Wnt signaling in colorectal cancer,reduce the incidence of colorectal cancer,and have a certain preventive effect.

To explore new teaching method of functional experiment,we opened the functional laboratory to encourage students' creativity and train their practical ability and develop their innovative ability.The experiment teaching innovation was attempted and discussed the functional speciality.

Objective To study the role of survivin in the incidence of pulmonary hypertension secondary to congenital heart disease (CHD) of left-to-right shunt. Methods Forty-nine cases were divided into 4 groups according to mean pulmonary artery pressure (mPAP), that is, non-hypertension group, mild hypertension group, medium hypertension group, and severe hypertension group. Their lung tissue samples were collected during heart surgery. TUNEL technique and immunohistochemical analysis were respectively used to detect nuclear DNA fragment of apoptotic cells and the expression of survivin, bcl-2 and bax. Western blotting also was conducted to detect survivin expression. Results The pulmonary wall of CHD patients with medium or severe pulmonary hypertension became significant incrassate. The apoptotic index and expression level of bax were significantly decreased but surviving and bcl-2 expressions were significantly increased in CHD with medium or severe pulmonary hypertension. Survivin expression was significant correlation with percentage media wall thickness and apoptotic index. Conclusion Apoptosis associated with survivin plays an important role in pulmonary artery remodeling after pulmonary hypertension of CHD.

Objective To summarize the effect of venous anastomisis from left atrium-common venous anastomisis ( supracardiac anasto-mosis) at the top of left atrium,and to find the best method to treat total anomalous pulmonary venous connection ( TAPVC) . Methods 52 cases,of which 35 male and 17 female with the age of 1 month to 41 years old and the weight of 3. 1~77 kg,hospitalized in West China hos-pital from January 2000 to April 2008,were treated by supracardiac anastomosis. Results One was dead and the other 51 cases were fully recovered and left hospital. After the operation,no anastomotic stenosis or arrhythmia was observed except the dead one. During follow-up peri-od which lasted from 3 months to 12 years,the heart function of 45 cases were normal. Conclusion supracardiac anastomosis can reduce the risk of anastomotic stenosis and arrhythmia,it is a promising method to treat supracardiac type TAPVC .

An efficient method for the analysis of multiclass plant growth regulators and pesticide (imidacloprid, acetamiprid) residues in tea was developed based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The samples were extracted with acetonitrile/formic acid (99∶1, V/V) solution, cleaned up with four sorbents including C18, strong anion exchanger (SAX), primary secondary amine (PSA) and anhydrous MgSO4. The compounds were separated on a HSS T3 column under positive/negative electrospray ionization mode, detected by scheduled multiple reaction monitoring (SMRM), and quantified by matrix-matched external standard curves. All pesticide residues showed good linearity in the concentration range of 1-200 μg/L (6-benzylaminopurine, paclobutrazol, uniconazole, forchlorfenuron, mepiquat chloride, imidacloprid, acetamiprid) or 5-1000 μg/L (2,4-dichlor-ophenoxyacetic acid, 4-chlorophenoxyacetic acid, indole-3-acetic acid, gibberellic acid, 1-naphthaleneacetic acid, indole-3-butyric acid) , with correlation coefficient (R2≥0.99). Limits of detection (LOD, S/N=3) and limits of quantitation (LOQ, S/N=10) were 0.18-9.68 μg/kg and 0.61-32.26 μg/kg, respectively. In addition, the spiked recoveries of tea samples were 73.1％-108.9％, and RSDs were 0.6％-8.0％. This method was applied to commercial samples, and all the detections were confirmed by acquiring transitions for each pesticide in the samples.

SUMOylation is a post‐translational modification involved in various cellular processes .SUMO‐specific protease (SENP) regulates SUMOylation by removing SUMO from conjugated substrates (deSUMOylation) and promoting maturation of SUMO precursor .In order to express Giardia lambia (C2 strain) SENP catalytic domain in E .coli ,the full‐length open reading frame of SENP was amplified by PCR from Giardia lamblia genome DNA .The PCR product about 1 620 bp was cloned into cloning vector pGM‐T .Sequencing result showed the sequence of SENP in C2 strain was identical with that in Gi‐ardia WB strain .Bioinformatics analysis showed that SENP protein possessed a 372 aa discontinuous ULP catalytic domain at C‐terminal .The catalytic domain of SENP was cloned into prokaryotic expression vector pET‐28a(+ ) .The recombinant vector pET‐28a(+ )‐SENPc was transformed into E .coli Rosetta(DE3) ,then the recombinant SENPc protein was expressed by IPTG induction .SDS‐PAGE and Western blot using anti‐His Tag antibody showed that the expression product of SENPc was a fusion protein with a molecular weight of 43 kD .The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SENP protein provide basis for further functional study of SENP .