Escherichia coli AFP111 is a spontaneous mutant with mutations in the glucose specific phosphotransferase system (ptsG) in NZN111 (delta pflAB deltaldhA). In AFP111, conversion of xylose to succinic acid generates 1.67 molecule of ATP per xylose. However, the strain needs 2.67 molecule ATP for xylose metabolism. Therefore, AFP111 cannot use xylose due to insufficient ATP under anaerobic condition. Through an atmospheric and room temperature plasma (ARTP) jet, we got a mutant strain named DC111 that could use xylose under anaerobic condition in M9 medium to produce succinic acid. After 72 h, DC111 consumed 10.52 g/L xylose to produce 6.46 g/L succinic acid, and the yield was 0.78 mol/mol. Furthermore, the reaction catalyzed by the ATP-generating PEP-carboxykinase (PCK) was enhanced. The specific activity of PCK was 19.33-fold higher in DC111 than that in AFP111, which made the strain have enough ATP to converse xylose to succinic acid.
Atmosphere ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Industrial Microbiology ; Metabolic Engineering ; Mutation ; Plasma Gases ; pharmacology ; Succinic Acid ; metabolism ; Temperature ; Xylose ; metabolism

Escherichia coli BA002, in which the ldhA and pflB genes are deleted, cannot utilize glucose anaerobically due to the inability to regenerate NAD+. To restore glucose utilization, overexpression of nicotinic acid phosphoribosyltransferase (NAPRTase) encoded by the pncB gene, a rate-limiting enzyme of NAD(H) synthesis pathway, resulted in a significant increase in cell mass and succinate production under anaerobic conditions. However, a high concentration of pyruvate was accumulated. Thus, co-expression of NAPRTase and the heterologous pyruvate carboxylase (PYC) of Lactococcus lactis subsp. cremoris NZ9000 in recombinant E. coli BA016 was investigated. Results in 3 L fermentor showed that OD600 is 4.64 and BA016 consumed 35.00 g/L glucose and produced 25.09 g/L succinate after 112 h under anaerobic conditions. Overexpression of pncB and pyc in BA016, the accumulation of pyruvic acid was further decreased, and the formation of succinic acid was further increased.
Anaerobiosis ; Escherichia coli ; enzymology ; genetics ; metabolism ; Fermentation ; Genetic Engineering ; Glucose ; metabolism ; Industrial Microbiology ; Lactococcus lactis ; enzymology ; NAD ; metabolism ; Pentosyltransferases ; biosynthesis ; genetics ; Pyruvate Carboxylase ; biosynthesis ; genetics ; Succinic Acid ; metabolism

Objective To study the expression of serum interleukin (IL )-6 and IL-10 in sepsis patients ,and their correlations with immune function .Methods A total of 56 patients with sepsis were divided into three groups ,including 18 patients in sepsis group ,21 patients in severe sepsis group ,17 patients in septic shock group .All the patients were also divided into survivor and death group according to whether they survived within 2 weeks .Other 30 healthy persons were selected in the control group .Serum IL-6 and IL-10 ,CD4+ /CD8+ were detected and compared .Results Serum IL-6 and IL-10 levels in the three sepsis groups were significantly higher than those in control group ,and with the severity aggravating ,these indicators increased .The level of serum IL-6 in sepsis patients was significantly reduced at the 3rd day ,while there was no difference on the serum IL-10 in severe sepsis group and septic shock group compared with that at admission(P>0 .05) .Compared with survivor group ,serum IL-6 and IL-10 levels on both admission and the 3rd in death groups increased significantly (P<0 .05) .CD4+ ,CD8+ ,CD4+ /CD8+ were negative related with serum IL-6 and IL-10 in both severe sepsis and septic shock group (P<0 .05) .Conclusion The expression of IL-6 and IL-10 in patients with sepsis are over-expressive ,and closely related with disease activity and immune function .

Objective To observe the curative effect of repairing the distal foot skin and soft tissue defect by using flap.Methods We used nutrient artery flap and low external ankle to repair the foot distal skin soft tissue defect in 16 cases.Results The flap survived,necrosis happened in small skin edge part and distal skin flap of sural nerve nutrient artery flap in two cases,and the skin graft healing with dots after removed the necrotic tissue and changed dressing.16 cases were followed up for 3-16 months,2 cases of flap slightly bloated,1 case back to the hospital got thin skin flap repairing technique;14 cases of walking were as usual,2 lame cases,related to some tissue defect in patients with forefoot injury.The use of flap to repair the foot injury,could maximially recover the limb function,reduce the rate of the sick.Conclusion Skin flap is a kind of operation which is simple,with smaller cost for skin area,beautiful and effective to repair skin and soft tissue defect of foot.

Objective To observe the clinical significance and effect of dysfunction of dendritic cell( DC) in colorectal cancer with liver metastasis.Methods Peripheral blood were respectively collected from healthy adult donors (30 cases), preoperative and postoperative coloreatal cancer patients without liver metastasis (30 cases) , and 30 postoperative coloreatal cancer patients with liver metastasis from Jan 2008 to Jun 2010.Peripheral blood mononuclear cells were separated, GM-CSF( 1000 U/ml) , IL-4( 1000 U/ml) and TNF-α ( 1000 U /ml) were added into cell culture fluid to induce the mononuclear cells for mature dendritic cells.There were two subgroups, and in antigen processing subgroup the lysate of HT29 colonic carcinoma cells (100 μg/ml) were added into the cell culture fluid.The T lymphocytes from healthy adults were added into two subgroups by ratio of 1∶10 ( DCs∶ T cells) , cocultured for 7 days.The level of INF-γ and IL-10 in cell culture fluid was assayed with ELISA method.The optical density (OD) of CCK8 ans LDH was assayed with ELIASA to indirectly measure the reproductive activity and the killing efficacy of T lymphocytes.Results The IL-10 level in cocultured fluid of peripheral blood DCs in postoperative colorectal carcinoma patients with liver metastasis and T lymphocytes of healthy adults was significantly higher than that of preoperative patients of colorectal carcinoma and health adults without tumor antigenic stimulation(11.9±1.3) pg/ml vs.(29.6±9.7) pg/ml, (23.4±8.0) pg/ml, F =4.475, P ＜0.05).The IFN-γ level in cocultured fluid of peripheral blood DCs in postoperative colorectal carcinoma patients with liver metastasis and T lymphocytes of healthy adults was significantly higher than that of postoperative patients of colorectal carcinoma and healthy adults with or without tumor antigenic stimulation ( 34 ± 9) pg/ml vs.(26 ± 12 ) pg/ml, (24 ± 6) pg/ml, F = 5.206, P ＜ 0.05).The killing activity of healthy adults T lymphocytes induced by HT29 colonic carcinoma cells in postoperative colorectal carcinoma patients with liver metastasis was significantly higher than that of preoperative patients of colorectal carcinoma and healthy adults (30.6 ±8.6) pg/ml vs.(12.1 ±2.4) pg/ml, (14.9 ± 1.7) pg/ml, F =4.147, P ＜ 0.05).Conclusions T lymphocytes produce IL-10 when indued by DCs from patients with colorectal carcinoma under stimulation of tumor antigen leading to tumor immune escape and liver metastasis.The killing activity of T lymphocytes can be enhanced when stimulated by exogenous tmuor antigen.

Inguinal incarcerated hernia in adults is a common acute abdomen in hernia and abdominal wall surgery. If not treated in time, it is easy to progress to constrictive hernia, lead to intestinal ischemic necrosis, cellulitis of tegmental tissue outside the hernia, hernia sac empyema, intestinal fistula, and even cause toxic shock, with significantly increased mortality. The types of incarceration are different and the corresponding management methods are different. Based on the proposal of the concept of musculopubic foramen hernia repair, inguinal incarcerated hernia includes incarcerated indirect hernia, incarcerated direct hernia, incarcerated femoral hernia, etc. At present, there is no uniform standard for the comprehensive treatment of different types of inguinal incarcerated hernia, and the clinical management strategies of adult inguinal incarcerated hernia still face serious challenges.

Objective:To investigate the safety and efficacy of drug-eluting stents(DES)in patients with acute coronary syndrome(ACS)complicated with diabetes and multivessel disease.Methods:A total of 80 patients with ACS who received XIENCE V Everolimus stent implantation in the heart center of the 82nd army hospital from January 2017 to October 2018 were retrospectively enrolled and analyzed.Patients were divided into the diabetic group(35 cases)and the non-diabetic group(45 cases)according to whether they were complicated with diabetes or mltivessel lesions.Stent implantation status within 12 months after PCI were compared between the groups.Results:(1)All stents were successfully implanted in the two groups.After coronary angiography re-examination within 12 months, there were no significant differences in the rate of stent restenosis(0.0% vs 2.2%), non-fatal myocardial infarction(2.9% vs.2.2%), or MACEs(2.9% vs.4.4%)between the diabetic and non-diabetic groups(all P>0.05). No patients died from cardiogenic causes in the two groups.(2)During the follow-up period, one patient was hospitalized for acute myocardial infarction in the diabetic group and one case died of an ascending aorta rupture. Conclusions:The safety and efficacy of XIENCE V DES in patients with diabetes and multivessel lesions are good, and the incidence of MACEs is low.

Objective:To investigate the effect of bone mesenchymal stem cells derived exosomes (BMSC-Exo) on improving hippocampal microangiogenesis, energy metabolism, and behaviors in depression mouse models.Methods:(1) Mouse bone marrow mesenchymal stem cells were isolated and cultured to extract BMSC-Exo; BMSC-Exo morphology was observed by transmission electron microscopy, BMSC-Exo particle diameter ranges were determined by Zetaview analyzer, and expressions of CD9 and CD63 in BMSC-Exo were detected by Western blotting. (2) Depression models were established in 2 mice by chronic unforeseeable mild stress (CUMS); 24 h after stereotaxic injection of phosphate buffer solution (PBS) or DiR labeled BMSC-Exo, BMSC-Exo uptake was detected by in vivo imaging system. (3) Thirty-six mice were randomly divided into control group, model group and BMSC-Exo group ( n=12); CUMS was used to establish depression models in the latter 2 groups; brain stereotaxic injection of 1 μL BMSC-Exo was given to mice in the BMSC-Exo group after modeling, and same amount of PBS was given to the control group; behaviors were observed by forced swimming test (FST), tail suspension test (TST) and open field test (OFT); hippocampal microvascular length and number were detected by alkaline phosphatase staining; energy metabolism in the hippocampus was detected by micro positron emission tomography/computed tomography (mPET/CT); glucose transporter 1 (GLUT1) expression in the hippocampus was detected by Western blotting. Results:(1) BMSC-Exo had a typical disk-like vesicle-like structure with particle size of (100.5±1.4) nm; Western blotting confirmed that CD9 and CD63 expressed in BMSC-Exo. (2) In vivo imaging showed no fluorescence in the brain and liver after PBS injection, but obvious local fluorescence after BMSC-Exo injection. (3) Compared with the control group, the model group and BMSC-Exo group had significantly longer rest time in FST and TST and shorter movement distance and time in the central region of OFT ( P<0.05); compared with the model group, BMSC-Exo group had significantly shorter rest time in FST and TST and longer movement distance and time in the central region of OFT ( P<0.05). Compared with the control group, the model group and BMSC-Exo group had significantly decreased standard uptake value (SUV) of regions of interest, microvascular length and number, and GLUT1 expression in the hippocampus ( P<0.05); compared with the model group, the BMSC-Exo group had significantly higher SUV, microvascular length and number, and GLUT1 expression in the hippocampus ( P<0.05). Positive correlations were noted between hippocampal microvascular length and SUV and between microvascular number and SUV in the 3 groups ( r=0.540, P<0.001; r=0.600, P<0.001). Conclusion:BMSC-Exo could promote microangiogenesis energy metabolism in the hippocampus to improve depression-like behaviors in depression mouse models.

Objective To comapre and analyze the differences and commonalities of expression profiles of serum exosomal microRNA between patients with thyroid nodules and healthy persons at different iodine levels,and then provide evidence for screening early diag-nostic markers of thyroid nodules at different iodine levels.Methods The peripheral blood samples from 10 patients with thyroid nod-ules and healthy volunteers at different iodine levels were collected.Their serum iodine levels were measured by the arsenic cerium cat-alytic spectrophotometry.Serum exosomal microRNA were extracted and the expression levels of microRNA were determined by the high-throughput sequencing technology.The differential target genes were predicted and further performed Gene ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.Results Compared with healthy volunteers,there were 6 downreg-ulated miRNAs in the patients with thyroid nodules at different iodine levels,namely miR-324-5p,miR-6511b-3p,miR-9903,miR-550a-3p,miR-5001-3p,and miR-3688-3p.Differentially expressed exosomal microRNA could regulate the MAPK signaling path-way,PI3K-AKT signaling pathway,VEGF signaling pathway,and NF-κB signaling pathway.Conclusion Six differentially expressed microRNAs is identified,which may serve as biological markers for the early diagnosis of thyroid nodules at different iodine levels.