Objective To construct,express and analysis the antitumor activity of anti-EGFR ScFv(HL-L-L),and analyze bioinformatics and characterize correlation spectroscopy.Methods EGFR ScFv(HL-L-L)gene was obtained by OVER LAPING technology and construction of prokaryotic expression vector pET-28a-c(+)-VH-L-L.After induced expression,target protein HL-L-L was purified by Ni column,and its tertiary structures was predicted with SWISS - MODEL;size distribution and zeta potential distribution of the target protein HL-L-L were characterized by NanoZSP;the inhibition effects of HL-L-L on breast cancer cell proliferation were studied with the method of pharmacology.Results Prokaryotic expression vector pET-28a-c(+)-VH-L-L was constructed successfully,high purity of protein HL-L-L was obtained,and successfully predicted its three-dimensional structure;the size was about 489.7 nm,and the Zeta potential was 38.1 ,negatively charged.It had a certain inhibition effect on MCF # 7 proliferation,compared with blank group,and the difference was significant(P<0.01 ).Conclusion This study successfully express all anthropogenic medicinal anti-EGFR ScFv(HL-L-L),which has significant inhibiting effect on breast cancer cell proliferation.It provides the foun dation for further development of small molecules of anti-EGFR antibody drug.

Objective To investigate the roles of ubiquitin-conjugating enzyme(UBC9)in HSCs activa-tion and liver fibrosis. Methods Western blot was used to analyze the expression of UBC9 under the stimulus of different concentrations of TGF-β1. The effective shRNA-targeting UBC9 gene was synthesized and HCSs were in-stantaneously transfected using lipofectamine method. Non-specific shRNA-transfected group cells and shRNA-tar-geting UBC9-transfected group cells were set up. The mRNA and protein levels of UBC9 were determined with Quantitative Real-Time PCR and Western blot. Western blot also used to examine the expression level of collagenⅠ,α-SMA and P-smad3 after transfection of UBC9 shRNA into HCSs and CCK-8 assay was used to detect cell proliferative capacity after transfection. Results UBC9 expression was significantly up-regulated in TGF-β1-treat-ed HSCs. Knockdown of UBC9 significantly inhibited TGF-β1-induced HSCs proliferation,as well as decreased the expression levels of a-SMA and collagen I. Furthermore ,knockdown of UBC9 attenuated the phosphorylation of Smad3 in the presence of TGF-β1. Conclusions UBC9 may function as a novel regulator to modulate HSC activa-tion,potentially by inhibiting the TGF-β1/Smad3 signaling pathway,which reveals novel mechanistic insights into the anti-fibrotic effect of UBC9.

Objective To observe the effect of the advanced glycation end products(AGEs)on the iNOS expression in cultured cardiac microvascular endothelial cells.Methods Cultured the cardiac microvascular endothelial cells in vitro.After AGEs of the different dose(50～200 mg/L)and different times(0～24 h)played the role on the cells,we determined the NO generation and iNOS protein expression of all groups.Results NO generation and iNOS protein expression increased with the AGEs-dose increasing and treatment time.The results were significantly different.Conclusion These results demonstrate that AGEs may induce iNOS to produce toxic NO in cardiac microvascular endothelial cells,then the diabetic cardiomyopathy occures.

Objective: To investigate the effect of chromium picolinate on egg production, egg quality, serum biochemical indices and immune response. Methods: A total of 120 49 w old Beijing Red commercial layers were randomly allotted to one of five treatments, and fed a basal corn soybean meal diet and the basal diet supplemented with 0.4,2.0,10.0 or 20.0 mg/kg Cr as chromium picolinate under heat stress for 30 days.Results: Cr increased average egg weight(P0.20) on egg yield, feed intake and feed/egg ratio. Added Cr at 0.4 ,2.0,10.0 mg/kg enhanced (P

The aim of the experimental teaching educational reform is to improve teaching quality of pathology and increase the ability of linking theory with practice.The educational reform wag carried out by analyzing clinical pathological cases and integrating basic preclinical medicine mode.Through the educational reform,the students were guided to shape a whole medical knowledge system with the center of organ pathological change,and know well the learning method of connecting each subject,and the teaching quality of pathology Was thus successfully improved.

Objective To evaluate the clinical effect of endoscopic sphineterotomy (EST) and endoscopic sphincterotomy associated with traditional Chinese medicine in the treatment of intrahepafic duct stones.Methods Eighty patients with common bile duet stones and intrahepatic duct stone were randomly divided into a treatment group and a control group.EST associated with traditional Chinese medicine was performed in the treatment group,and EST only was used in the control group.Clinical effects were observed.Results Common bile Duet stone clearance was achieved in 94% patients in the treatment group,contrasting 88% in the control group.There was no significant difference between them (P＞0.05).While intrahepatic duct stone clearance was 74% and 28% respectively in the treatment group and the control group,showing significant difference (P＜0.05).Conclusion It is a good way that endoscopic sphincterotomy associated with traditional Chinese medicine treating intrahepatic duct stones.

Objective To explore the effects of gross saponins of Tribulus terrestris L.on inflammatory reaction and permeability of blood-brain barrier in rats following cerebral ischemia-reperfusion injury and their potential mechanisms. Methods Sixty SD rats were divided into sham operation group,model control group,gross saponins of Tribulus terrestris L.at low-dose (10 mg?kg-1 )and high-dose groups(30 mg?kg-1 ).Cerebral ischemia -reperfusion model was established with suture emboli method in middle cerebral artery of rats.Neural injury scores,the contents of Evans blue ( EB) and myeloperoxidase( MPO) activities in rat brain were measured 24 hours after the cerebral reperfusion post 2 h ischemia.Content of tumor necrosis factor-α (TNF-α) in rat brain was detected by ELISA; expression levels of matrix metalloproteinase-9(MMP-9) in rat brain was determined by Western blot. Results Compared to the model control group,the neurological deficit scores were significantly decreased(P<0.05),MPO activities and EB contents decreased(P< 0.05 or P< 0.01) in the treatment groups.The expression levels of TNF-α were significantly lower in the treatment groups(0.760±0.110) mg?g-1 and (0.670±0.073) mg?g-1 compared to (0.920±0.128) mg?g-1 in the model control group ( P< 0.05 or P< 0.01). The MMP-9 expression levels were (1.770± 0.181)% and(1.480±0.146)%,significantly lower than(2.200±0.186)% in the model control group(P<0.01). Conclusion Gross saponins of Tribulus terrestris L. exert neuroprotective effects on cerebral ischemia-reperfusion injury in rats through inhibiting the inflammatory reaction and decreasing the permeability of blood-brain barrier,which may be associated with the decrease of the TNF-α content and downregulation of the MMP-9 expression.

Objective To investigate the impact of preload fasting and meal replacement in patients with metabolic syndrome.Methods A total of 92 subjects with metabolic syndrome were enrolled in the study.They were assigned into the preload fasting group (PFG),the meal replacement group (MRG),and the control group (CG) for 12-weeks intervention.Special dietary with 100 kcal was provided 30 min before each meal in the PFG,and while in the MRG the same dietary was taken just before each meal and the amount of meal was reduced appropriately.The subjects in CG took meals as usual.Body mass index,waist circumference,and insulin resistance were assessed.Satiety situation was investigated by the scale.Results After 12 weeks,improvement were found in fasting insulin(-3.29 mU/L) and waist circumference (-4.04 cm) in the PFG and significant difference was shown compared to the CG (P＜0.05).Satiety index in the PFG was the most significant among the three group.Conclusion Preload fasting is helpful in improving insulin resistance,reducing waist circumference,and enhancing satiety.

Objective To explore the mechanism that gold nanorods trigger apoptosis in cancer cells.Methods Gold nanorods was synthesized by gold seed growing method, and its characterization was detected; gold nanorods on cell proliferation-toxicity were evaluated by CCK-8 Kit and apoptosis were detected by flow; mitochondrial membrane potential were tested by JC-1 and activation of Caspase 9 and Caspase 3 were detected by western blot. Results The results found that gold nanorods had nontoxic to normal cells, but highly toxic to tumor cells; and with the increasing of gold nanorods’ working time, the percentage of apoptotic cancer cells was increasing; in addition to, normal cells’ mitochondrial membrane potential did not change, but cancer cells had a significant reduction in mitochondrial membrane potential.Conclusion This study proves that gold nanorods induce apoptosis through the mitochondrial apoptosis pathway.