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Synthetic Biology is one of the most promising fields of modern Biology and a frontier interdisciplinary subject in the 21st century. With the rapid development of synthetic biology, the International Genetically Engineered Machine (iGEM) competition has emerged. The iGEM competition, based on the subject foundation of Synthetic Biology, intends to solve the biological problems in our daily life by applying modern biological technology. In recent years, with the continuous increase of participating teams, the iGEM competition has received extensive attention and achieved great progress. On the basis of the development of Synthetic Biology, we analyzed the 2018-2020 award-winning projects of the iGEM competition and illustrated the role and significance of the iGEM competition in cultivating college students' innovative thinking and ability with the participation experience of the iGEM team of Southwest Jiaotong University as an example.
Assuntos
Humanos , Engenharia Genética , Estudantes , Biologia Sintética , UniversidadesRESUMO
Objective To investigate the effect of different antibodies on Toll-like Receptor 4-High Mobility Group Box 1 and its downstream signal transductions in distant organ injuries caused by intestinal ischemia/reperfusion in mice.Methods A total of 40 mice (C57BL/6,SPF level) were by random number table method assigned into five groups:sham,control,anti-HMGB1,anti-Myeloid differentitation gene,and antiTIR domain containing adaptor inducing IFN-β (n=8).In the control,anti-HMGB1,anti-MyD88,and antiTRIF groups,the IgG,HMGB1,MyD88,and TRIF antibodies were injected,respectively,via the tail vein 30 minutes before ischemia (1 mg/kg body weight,0.025％).After anesthesia and abdomen incision,all mice,except the sham group,underwent intestinal ischemia by clamping the superior mesenteric artery for 60 minutes followed by 60 minutes of reperfusion.Sham group underwent the same surgical procedures except for clamping the artery.Serum nuclear factor-κB p65,Interleukin-6 and Tumor Necrosis Factor-α were measured.Morphological changes in the lung and intestine were evaluated.mRNA and protein expressions of HMGB1 and NF-κB in lung and intestinal tissues were assayed.Results Compared with the control group [(228.53± 24.85),(104.91±31.18),and (70.81±46.97) ng/L],HMGB1 [(145.00±33.63),(62.28±6.73),and (52.76± 5.71) ng/L],MyD88 [(191.12± 13.22),(85.90± 17.37),and (63.19 ± 5.47) ng/L],and TRIF [(183.73±10.81),(78.14±7.38),and (59.70±4.63) ng/L] significantly decreased the serum level of NF-κB (P=0.000,0.005,0.001),IL-6 (P=0.000,0.004,0.000) and TNF-α (P=0.000,0.024,0.002) after ischemia reperfusion.Tissue injuries in the lung and intestine were also alleviated by HMGB1,MyD88,and TRIF.The anti-HMGB1,anti-MyD88,and anti-TRIF groups displayed significant elevations of HMGB1 mRNA [lung (1.89±0.18),(2.35±0.31),and (2.29±0.28),ileum (4.93±0.55),(5.96± 0.73),and (5.76±0.51)],NF-κB mRNA [lung (1.42±0.23),(1.77±0.18) and (1.70±0.13),ileum (2.23±0.55),(3.11±0.38) and (2.99±0.24)] and NF-κB protein expressions in lung and ileum tissues compared to the sham group [lung HMGB1 mRNA (1.04±0.19) (P=0.000,0.000,0.000),NF-κBmRNA (1.03±0.21) (P=0.004,0.000,0.000),ileum HMGB1 mRNA (1.14±0.54) (P=0.000,0.000,0.000),NF-κB mRNA (1.03±0.23) (P=0.000,0.000,0.000)].However,incornparison with the control group [lung HMGB1 mRNA (2.67±0.23) (P=0.000,0.035,0.016),NF-κB mRNA (2.04±0.29) (P=0.000,0.039,0.012),ileum HMGB1 mRNA (6.70±0.66) (P=0.001,0.038,0.015),NF-κBmRNA (3.71±0.53) (P=0.000,0.018,0.006)],the other three groups showed a significant down-regulation,with the most remarkable decrement in the anti-HMGB1 group.Application of anti-HMGB1,anti-MyD88,and anti-TRIF could drastically attenuate the tissue injuries in ischemia reperfusion.anti-HMGB1 exhibited the most significant effect.Conclusions HMGB1 and its downstream signals play an important role in intestinal ischemia reperfusion injuries in mice.Of two downstream signals,the TRIF-dependent pathway exerts a more important effect than that of the MyD88-dependent pathway.
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Objective To investigate the effects of lymph from ischemic/reperfused intestine on the inflammatory factors and Toll-like receptor 4 (TLR4) ligand high mobility group box-1 (HMGB1) in TLR4 deficient (TLR4-/-) mice.Methods A total of 20 SD rats weighing (300 ±20) g were randomly assigned into two groups:lymph drainage group (group N,lymph drainage for 180 minutes without other treatment) and intestinal ischemia/reperfusion group (group I/R,draining the lymph for 180 minutes while clipping the superiormesenteric artery for 60 minutes followed by 120-minute reperfusion).Thirty-two TLR4-/-mice and thirty-two C57BL/6 wild type (WT) mice were each divided into 4 sub-groups (n =8),injected with different fluids through the caudal vein:group N with normal lymph;group I/R with I/R lymph;group Edt with endotoxin;group HMGB1 with HMGB1 protein.The mice were sacrificed 180 minutes after the injection for sample collection.Results The levels of endotoxin and HMGB1 in the lymph drainage of the group I/R rats were significantly higher than that of the group N rats [(0.034 ± 0.050) Eu/ml vs.(0.017 ± 0.023) Eu/ml,P =0.033;(4.293 ± 0.883) ng/ml vs.(0.509 ± 0.128) ng/ml,P =0.006].In the mice injected with HMGB1,the mucosa thickness and villus height in the ileum of the WT mice were significantly lower than that of the TLR4-/-mice [(335.8±43.2) μmvs.(602.1±37.5) μm,P=0.000;(273.0±31.7) μm vs.(404.5 ± 18.6) μm,P =0.000];in both WT and TLR4-/-mice injected with the I/R lymph drainage,the mucosa thickness and virus height were decreased,but the decrements were significantly lower in TLR4-/-mice;there were no statistically significant differences in the levels of interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),endotoxin,and HMGB1 between the TLR4-/-and the WT mice injected with normal lymph or endotoxin.In the mice injected with I/R lymph drainage,the levels of inflammatory factors in the TLR4-/-mice were significantly lower than those in the WT mice [TNF-α:(28.637 ±5.166) pg/ml vs.(41.917 ±8.175) pg/ml,P=0.000;IL-6:(60.900 ±24.729) pg/ml vs.(110.265 ±28.545) pg/ml,P =0.000].In the mice injected with HMGB1,the levels of inflammatory factors in the TLR4-/-mice were significantly decreased compared with those in the WT mice [TNF-α:(20.865 ± 6.464) pg/ml vs.(31.059 ± 6.204) pg/ml,P=0.004;IL-6:(36.268 ±8.977) pg/ml vs.(76.677 ± 14.099) pg/ml,P=0.000].Conclusions The concentrations of endotoxin and HMGB1 are significantly increased during intestinal I/R in rats.After injection of I/R lymph drainage,endotoxin,and HMGB1,the levels of inflammatory factors and HMGB1 in the mice injected with I/R lymph drainage are significantly higher than those in the mice injected with normal lymph;the levels of inflammatory factors and local damage of intestinal mucosa are significantly reduced in the TLR4-/-mice than in the WT mice.The gut-lymph pathway may play a key role in the intestinal I/R injury.
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Objective To evaluate the effects of ambroxol on prevention of bronchopulmonary complications after thoracotomy in patients with chronic obstructive pulmonary disease(COPD).MethodsTotal 161 elderly patients aged ≥70 years with COPD undergoing thoracotomy were randomly allocated to two groups:ambroxol group (300 mg/d,6 d) and placebo group as control.Pulmonary complications were evaluated by clinical symptoms,radiographic changes,and blood gas analysis. Results The incidence rates of atelectasis were 8.6％ in ambroxol group and 28.8％ in placebo group,respectively.The PaO2 values after surgery in ambroxol group decreased more than in placebo group (P＜0.05) compared with the preoperative values.Side effects were not found in all patients.Conclusions Ambroxol should be considered as an alternative pharmacologic approach for the prevention of post-thoracotomy pulmonary complications in the elderly patients with COPD.