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 study the effect of LDYS-14007 on JAK1-STAT3 signaling pathways.Methods MDA-MB-231 cells were treated with 10μmol,1 nmol LDYS-14007, and 10 μmol Tofacitinib,respectively.Western blot assay was used to determine the expression of JAK1,Phospho-JAK1, STAT3 and Phospho-STAT3.Results The absorbance value was linearly related to the concentration of protein C, The linear equation is A=0.0075C+0.0029, r=0.9976, The linear range of 1.08-5.08 mg/mL, With the increased concentration of LDYS-14007, the amount of Phospho-JAK1, Phospho-STAT3 were all gradually decreased.Conclusion LDYS-14007 leads to the levels of Phospho-JAK1 and Phospho-STAT3 decrease, which inhibits JAK1-STAT3 signaling pathway.LDYS-14007 may play an important role in the treatment of rheumatoid arthritis.

Objective To investigate the impacts of prehospital endotracheal intubation on postoperative pulmonary infection in the patients with severe traumatic brain injury.Methods Retrospectively, the clinical data of 284 patients with severe traumatic brain injury admitted in the 97th Hospital of PLA from July 2007 to December 2012 were analyzed.The patients were classified into two groups according to the timing of endotracheal intubation, namely, prehospital intubation group and admission intubation group.The postoperative pulmonary infection incidence, occurrence time, and the duration of treatment of the two groups were studied.Results The incidences of postoperative pulmonary infection in patients intubated before and after admission were 38.0％ and 25.2％ respectively.Pulmonary infection occurred in the prehospital intubation group was at the (9.9 ± 0.6) d after admission, and in the admission intubation group was at the (11.6 ± 0.3) d after admission.The duration of treatment for postoperative pulmonary infection was (21.2 ± 7.2) days and (14.5 ± 9.0) days respectively.Compared with the patients intubated after admission, patients intubated before admission suffered higher incidence (P ＜ 0.05), earlier onset (P ＜ 0.05), and longer treatment duration of pulmonary infection (P ＜ 0.01).Conclusions Severe traumatic brain injury patients with prehospital endotracheal intubation are more susceptible to pulmonary infection.Avoiding the tracheal injury and bacterial contamination in the procedure could reduce the incidence of pulmonary infection.

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.

In order to improve the expression of recombinant human atrial natriuretic peptide (ANP), a new plasmid (pET28a(+)/ANP₃) containing 3 tandem ANP genes with lysine codon as the interval linker, was constructed. Target gene was transformed into Escherichia coli BL21 (DE3) and induced by IPTG, about 60% of the total-cell-protein was the target protein, His₆-ANP₃. After denaturation and refolding, it was digested by Endoproteinase Lys-C and Carboxypeptidase B (CPB) and then purified by a series of purification processes, about 16 mg purified ANP monomer could be obtained from one liter bacteria broth of shaking culture. Ultimately, the purity of protein was above 90% determined by UPLC and Tricine SDS-PAGE, its molecular weight was 3 080 Da according to LC-MS identification and it was proved to be equivalent to the reference product by ELISA. The use of tandem gene expression can provide a new possible model for the expression of other peptide drugs.
Atrial Natriuretic Factor ; biosynthesis ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; metabolism ; Gene Expression ; Humans ; Metalloendopeptidases ; Peptides ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis