Heme peroxidase EfeB in E. coli belongs to the dye-decolorizing peroxidase (DyP) superfamily. Peroxidases in this superfamily have a good ability in degradation of synthetic dyes, but their physiological functions in organisms are unclear. In order to further understand the physiological function of EfeB, the mutant strain EcoΔefeB was constructed by homologous recombination. The differences between parental strain E. coli BL21 and EcoΔefeB at genome transcription level as well as cell growth under different conditions were compared. The response of efeB to iron ion was also investigated. The results showed that the deletion of efeB gene caused the differential expression of 1 765 genes, which were mainly related to cell metabolic pathway, cell membrane synthesis and flagellum movement. There was no significant difference in cell growth between BL21 and EcoΔefeB at pH 7. 0, but the growth of BL21 was much better than that of EcoΔefeB at pH 4. 5. The functional expression of efeB may support the survival of E. coli at low pH. EfeB was significantly up-regulated when Fe

Objective To compare the clinical efficacy and safety of two different conditioning regimens in nonmyeloablative autologous peripheral blood stem cell transplantation (NAST) for the treatment of systemic lupus erythematosus (SLE).Methods Different conditioning regimens were used in two groups:cytarabin combined cyclophosphamide in group 1 and ATG combined cyclophosphamide in group 2.Different recovery time of leucocytes,neutrophils and platelets in the two groups were compared.Statistical analysis were carried out by paired t-test.Results The mean time for peripheral leucocytes reaching 1.0×109/L,neutrophils getting up to 0.5×109/L,platelet raising to 100×l09/L and hemoglobin rising to 120 g/L in group 1 were [(7.2±1.3),(8.0±1.5),(10.5±1.4),(22.1±2.3)days] and [(10.4±2.1),(12.0±1.9),(19.3±2.1),(28.1± 2.4)] days in group 2.The difference was statistically significant (P＜0.01).CD4+ cell count and the ratio of CD4+/CD8+ of pre- and pro-NAST was changed.No significant differences were observed in the two groups.Conclusion For the sake of safety and hematopoietic reconstitution,we recommend cytarabin combined cyclophosphamide as the preferred conditioning regimen.

Objective To investigate the long-term efficacy of nonmyeloablative autologous peripheral blood stem cell transplantation(NAST) to cure refractory autoimmune disease(AD).MethodLong-term follow up of four cases of AD patients with NAST were summarized.The pretreatment regimen was intravenous injection of cytarabin (200 mg· kg-1· d-1 ) and cyclophosphamide (40 mg· kg-1· d-1).The therapeutic effect was evaluated by the change of symptoms and signs and long term complications.Changes of immune function were detected by flow-cytometry.ResultsFive cases of patients had been successfully engrafted.The average time for peripheral leucocytes count to reach 4.0×109/L was 12 days.It needed 10 days for platelets to return to 100×109/L and 22 days for hemoglobin to 120 g/L.Apparent remission of symptoms and signs was observed after transplantation.Lymphocyte subtypes analysis pre- and post- NAST showed that count of CD4+ and the ratio of CD4 +/CD8 + was returned to normal.One patient gave birth to a healthy baby four years after transplantation.Three female patients returned tonormal life. Conclusions Compared with classical myeloablative stem cell transplantation,NAST has a rapid hematopoietic recovery and good long-term therapeutic effect in AD.The quality of life in AD patients treated with NAST is higher than those treated with myeloablative hematopoietic stem cell transplantation.

Objective: To investigate the effect of menin protein encoded by the multiple endocrine neoplasia type I (MEN 1) gene on proliferation of human gastric cancer AGS cells and its possible machanism. Methods: An adenovirus encoding MEN 1 gene was used to infect human gastric cancer AGS cells. The expression level of menin protein in AGS cells infected with recombinant adenovirus Ad-MEN1 was detected by Western blotting and immunofluorescence. The proliferation of AGS cells was tested by MTT method. The cell cycle distribution and apoptosis of AGS cells were analyzed by flow cytometry (FCM). The expressions of phospho-Akt (p-Akt) and nuclear factor-kappa B (NF-κB) which were related to phosphoinositide 3-kinase (PI3K)-protein kinase B (PKB) signal pathway were detected by Western blotting. Results: The results of Western blotting and immunofluorescence showed that the expression of menin protein in gastric cancer cell line AGS was increased after Ad-MEN1 infection (P < 0.05). The infective efficiency of recombinant adenovirus Ad-MEN1 was the highest when the multiplicity of infection was 60 in AGS cells, and the cell proliferation was obviously inhibited (P < 0.05). The percentage of cells was increased in phase G0/G1 and decreased in phase S (P < 0.05). However, the apoptosis rate of AGS cells after infection with Ad-MEN1 was not significantly changed (P > 0.05). The expressions of p-Akt and NF-κB p65 in AGS cells were decreased after the expression of menin in AGS cells was increased (P < 0.05). Conclusion: This study reveals that over-expression of menin can inhibit the growth of human gastric cancer cell line AGS. This machanism may be related to the inhibition of PI3K/Akt and NF-κB signal transduction pathways.

Intein is a part of polypeptide in the premature protein with the capability of self-splicing, which is widely applied in protein purification, protein conjuction, cyclopeptide preparation, protein labeling and biosensor. In this review, we summarized the development of intein used in protein purification, discussed intein-mediated chromatographic and non-chromatographic purification systems, and summarized the researches in manipulating intein cleavage reaction. This work is to provide clues for improvement of intein-mediated protein purification.
Chromatography, Affinity ; Inteins ; Protein Splicing ; Proteins ; isolation & purification

Objective:To explore the effect of somatosensory interactive games on lower-limb function after stroke. Methods:Randomized controlled trials (RCT) about the effectiveness of somatosensory interactive games on lower-limb function after stroke were retrieved from domestic and foreign databases, from inception to September, 2018. After literature quality evaluation, a meta-analysis was performed using RevMan5.3. Results:A total of 414 patients were included in eleven articles. Compared with routine rehabilitation measures, somatosensory interactive games improved Berg Balance Scale scores (WMD = 1.75, 95%CI 0.95~2.54, P < 0.001), increased stride frequency (SMD = 1.21, 95%CI 0.03~2.38, P = 0.04), decreased the time of Timed Up and Go Test (WMD = -4.21, 95%CI -7.36~-0.89, P = 0.01), and improved the motor function of lower limbs (SMD = 0.66, 95%CI 0.15~1.17, P = 0.01), but worked less in pace (SMD = 0.05, 95%CI -0.98~1.09, P = 0.92) and 10-metre Maximum Walking Speed (WMD = 3.54, 95%CI -1.12~8.20, P = 0.14). Conclusion:Compared with the routine rehabilitation, somatosensory interactive games can improve balance, walking and motor function. However, it is needed to further research on the pace and speed.

NAD kinase catalyzes the phosphorylation of coenzyme I [NAD(H)] to form coenzyme II [NADP(H)], and NADPH is an important cofactor in L-isoleucine biosynthesis. In order to improve NADPH supply, ppnK, the gene encoding NAD kinase in Corynebacterium glutamicum was cloned and separately expressed in an L-isoleucine synthetic strain, Brevibacterium lactofermentum JHI3-156, by an inducible expression vector pDXW-8 and a constitutive expression vector pDXW-9. Compared with the control strain JHI3-156/pDXW-8, NAD kinase activity of the inducible ppnK-expressing strain JHI3-156/pDXW-8-ppnK was increased by 83.5%. NADP(H)/NAD(H) ratio was also increased by 63.8%. L-isoleucine biosynthesis was improved by 82.9%. Compared with the control strain JHI3-156/pDXW-9, NAD kinase activity of the constitutive ppnK-expressing strain JHI3-156/pDXW-9-ppnK was increased by 220%. NADP(H)/ NAD(H) ratio and NADPH concentration were increased by 134% and 21.7%, respectively. L-isoleucine biosynthesis was increased by 41.7%. These results demonstrate that NAD kinase can improve the coenzyme II supply and L-isoleucine biosynthesis, which would also be useful for biosynthesis of other amino acids.
Brevibacterium ; genetics ; metabolism ; Cloning, Molecular ; Corynebacterium glutamicum ; enzymology ; Isoleucine ; biosynthesis ; Metabolic Engineering ; NAD ; metabolism ; NADP ; metabolism ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

Background: Phototherapy is an important method to treatvitiligo. However, it is unclear how phototherapy affectsmelanocyte precursors and skin neural crest stem cells. Objective: To investigate the underlying mechanisms of narrow-band ultraviolet B (NB-UVB) induced melanocyte lineagedifferentiated from human scalp-derived neural creststem cells (HS-NCSCs). Methods: HS-NCSCs were expandedfrom scalp hair follicles. The c-Kit−/CD57− HS-NCSCs wereisolated by cell sorting. Different doses of NB-UVB wereused to irradiate these HS-NCSCs. Cell ultrastructure was examinedby transmission electron microscope. Melanocytemarker expression was analyzed by Quantitative RT-PCRand Western blot. Cell proliferation and migration were alsoevaluated. Results: The c-Kit−/CD57− HS-NCSCs expressedembryonic NCSC biomarkers. NB-UVB at a dose of 100 mJof NB-UVB had little effect on the cell proliferation of differentiatedmelanocytes from c-Kit−/CD57− HS-NCSCs, while700 mJ inhibited cell proliferation significantly. The dendriticprocesses of differentiated melanocytes increased afterradiation. The tyrosinase and Melanocortin 1 receptor (Mc1R)expression of differentiated melanocytes increased after NB-UVB exposure. The effect of NB-UVB on tyrosinase expressionwas modulated by signaling inhibitors H89 andPD98059 as well as Mc1R level in the cells. The migrationability of differentiated melanocytes was enhanced under100 mJ exposure. Conclusion These data demonstrate thatNB-UVB facilitates melanocytic differentiation of the HSNCSCsand enhances migration of these cells. Mc1R andcAMP pathway play a critical role in NB-UVB induced melanocyticdifferentiation.

BACKGROUNDBiliverdin (BV) has a protective role against ischemia-reperfusion injury (IRI). However, the protective role and potential mechanisms of BV on lung IRI (LIRI) remain to be elucidated. Thus, we aimed to investigate the protective role and potential mechanisms of BV on LIRI.

METHODSLungs were isolated from Sprague-Dawley rats to establish an ex vivo LIRI model. After an initial 15 min stabilization period, the isolated lungs were subjected to ischemia for 60 min, followed by 90 min of reperfusion with or without BV treatment.

RESULTSLungs in the I/R group exhibited significant decrease in tidal volume (1.44 ± 0.23 ml/min in I/R group vs. 2.41 ± 0.31 ml/min in sham group; P< 0.001), lung compliance (0.27 ± 0.06 ml/cmH2O in I/R group vs. 0.44 ± 0.09 ml/cmH2O in sham group; P< 0.001; 1 cmH2O=0.098 kPa), and oxygen partial pressure (PaO2) levels (64.12 ± 12 mmHg in I/R group vs. 114 ± 8.0 mmHg in sham group; P< 0.001; 1 mmHg = 0.133 kPa). In contrast, these parameters in the BV group (2.27 ± 0.37 ml/min of tidal volume, 0.41 ± 0.10 ml/cmH2O of compliance, and 98.7 ± 9.7 mmHg of PaO2) were significantly higher compared with the I/R group (P = 0.004, P< 0.001, and P< 0.001, respectively). Compared to the I/R group, the contents of superoxide dismutase were significantly higher (47.07 ± 7.91 U/mg protein vs. 33.84 ± 10.15 U/mg protein; P = 0.005) while the wet/dry weight ratio (P < 0.01), methane dicarboxylic aldehyde (1.92 ± 0.25 nmol/mg protein vs. 2.67 ± 0.46 nmol/mg protein; P< 0.001), and adenosine triphosphate contents (297.05 ± 47.45 nmol/mg protein vs. 208.09 ± 29.11 nmol/mg protein; P = 0.005) were markedly lower in BV-treated lungs. Histological analysis revealed that BV alleviated LIRI. Furthermore, the expression of inflammatory cytokines (interleukin-1β, interleukin-6, and tumor necrosis factor-β) was downregulated and the expression of cyclooxygenase-2, inducible nitric oxide synthase, and Jun N-terminal kinase was significantly reduced in BV group (all P< 0.01 compared to I/R group). Finally, the apoptosis index in the BV group was significantly decreased (P < 0.01 compared to I/R group).

CONCLUSIONBV protects lung IRI through its antioxidative, anti-inflammatory, and anti-apoptotic effects.

Interleukin-33 (IL-33) is a multifunctional gene in the interleukin-1 family and is widely expressed in human tissues. Its receptor is ST2, is associated with asthma, diabetes, Alzheimer's disease, cardiovascular disease, inflammatory bowel disease, and cancer. IL-33, enhancing lipid metabolism and down-regulating adipogenic gene expression by inducing Th2 cytokine production, plays a protective role in fatty liver. In hepatitis, the IL-33/NKT/IFN-γ pathway exacerbates mouse hepatitis. While, the IL-33/ILC2/IL-13/STAT6 pathway reduces liver damage. The IL-33/ILC2/IL-13/IL-4Rα/STAT6 axis plays a facilitating role in cirrhosis. In liver cancer, IL-33/ILC2/IL-13 signaling pathway is found to promote cholangiocarcinoma in a favorable pathological environment, and hepatocellular carcinoma occurs by IL-33/IL-6/JAK/STAT/Mcl-1 signaling pathway.