A total of 256 medical professionals at the hospitals of different levels were interviewed and requested to complete the relevant questionnaires.The findings revealed that the top five unsatisfied items were growth & development,working conditions,doctor-patient relationship,income and social status.The levels of job satisfaction varied with their specific occupations and technical titles.Around two thirds of them once had the thought of leaving their pre-hospital care posts.

Objective To investigate the anti-apoptosis effect of transgiutaminase 2 (TG2)in osteosarcoma cell line MG-63 and to explore the mechanism of inhibiting apoptosis of tumor cells.Methods The TG2-tgrgeted siRNA was designed,and the hypoxia culture model of MG-63 cells was established by a hypoxia incubator and the cells were divided into four groups:normal oxygen group,the cells were cultured under normal oxygen;hypoxia group, the cells were cultured in hypoxic incubators;control siRNA hypoxia group,the cells were cultured in hypoxic incubators after transfection of control siRNA;TG2 siRNA hypoxia group , the cells were cultured in hypoxic incubators after transfection of TG2 siRNA.The expressions of Bax and cytochrome C (Cyt C)and the apoptotic rates were observed at different time (6,12,24,48,and 72 h)after hypoxia culture.Microtiter plate assay was performed to monitor the intracellular TG2 activity.RT-PCR was used to detect the mRNA expressions of TG2 and Bax.The expressions levels of TG2,Bax and Cyt C were observed by immunohistochemical staining and Western blotting method.The apoptotic rates were analyzed using flow cytometry.Results Compared with normal oxygen group,the activity of TG2,the mRNA and protein expression levels of TG2 in hypoxia group and control siRNA hypoxia group were increased remarkably with the prolongation of the hypoxia time (P<0.01);the expression level of Bax protein was decreased significantly (P<0.01 ), but the expression level of Bax mRNA had no significant change (P>0.05);the expression levels of Cyt C protein in cytoplasm and mitochondria and the apoptotic rates had no markedly changes (P>0.05).Compared with hypoxia group and control siRNA hypoxia group,the expression levels of mRNA and protein of TG2 in TG2 siRNA hypoxia group were decreased significantly at different time points (P<0.01);the protein expression levels of Bax and Cyt C in cytoplasm and the apoptotic rates were increased markedly (P<0.01 );the expression level of Cyt C in mitochondria was decreased (P<0.01).Conclusion TG2 can inhibit the apoptosis of tumor cells through down-regulating the Bax expression and preventing Cyt C releasing into the cytoplasm.

Carboxyl-terminal processing protease of D1 protein (CtpA) catalyzes carboxyl terminal processing of the D1 protein of photosystem II, which is essential for the assembly of a manganese cluster and consequent light-mediated water oxidation. It is a target for the discovery of wide-spectrum herbicide. We amplified the CtpA gene from spinach cDNA with standard PCR method and constructed it into pET-28a vector to generate a recombinant expression plasmid. Recombinant CtpA fusion protein with His-tag was expressed as soluble protein in Escherichia coli BL21(DE3) after induction with 0.1 mmol/L IPTG at 8 degrees C for 72 h. We purified the CtpA protein with the Ni-NTA affinity chromatography and Superdex 75 gel filtration chromatography respectively, and verified the protein by SDS-PAGE and Western blotting with anti-his antibody. Hydrolysis activity of CtpA was assayed by HPLC method with a synthetic 24-mer oligopeptide corresponding to carboxyl terminal of precursor D1 protein, and gave a total activity of 1.10 nmol/(mg x min). We used the purified CtpA protein as antigen to immune rabbit for the production of polyclonal antibody, and prepared antibody with high specificity and sensitivity. The results obtained in this paper provided the feasibility of high-throughput screening of lead compounds for the protease as inhibitors and mechanism analysis of CtpA enzyme.
Algal Proteins ; Antibodies ; metabolism ; Carboxypeptidases ; biosynthesis ; chemistry ; genetics ; immunology ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Hydrolysis ; Proprotein Convertases ; biosynthesis ; chemistry ; genetics ; immunology ; RNA, Plant ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Spinacia oleracea ; enzymology ; genetics

The mesencephalic astrocyte-derived neurotrophic factor (MANF) has been recently identified as a neurotrophic factor, but its role in hepatic fibrosis is unknown. Here, we found that MANF was upregulated in the fibrotic liver tissues of the patients with chronic liver diseases and of mice treated with CCl₄. MANF deficiency in either hepatocytes or hepatic mono-macrophages, particularly in hepatic mono-macrophages, clearly exacerbated hepatic fibrosis. Myeloid-specific MANF knockout increased the population of hepatic Ly6C^high macrophages and promoted HSCs activation. Furthermore, MANF-sufficient macrophages (from WT mice) transfusion ameliorated CCl₄-induced hepatic fibrosis in myeloid cells-specific MANF knockout (MKO) mice. Mechanistically, MANF interacted with S100A8 to competitively block S100A8/A9 heterodimer formation and inhibited S100A8/A9-mediated TLR4-NF-κB signal activation. Pharmacologically, systemic administration of recombinant human MANF significantly alleviated CCl₄-induced hepatic fibrosis in both WT and hepatocytes-specific MANF knockout (HKO) mice. This study reveals a mechanism by which MANF targets S100A8/A9-TLR4 as a "brake" on the upstream of NF-κB pathway, which exerts an impact on macrophage differentiation and shed light on hepatic fibrosis treatment.