Effects of exogenous carbon monoxide-releasing molecules 2 on the vitality and toxicity of E.coli / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on the vitality and toxicity of E. coli ATCC 25922, and to analyze the potential mechanism.</p><p><b>METHODS</b>(1) In vitro experiments. Standard strains of E. coli ATCC 25922 were divided into groups A (without addition), B, C, D, and E according to the random number table, and then the latter 4 groups were respectively cultured with 1.2 mmol/L CORM-2, 1.6 mmol/L CORM-2, 1.2 mmol/L inactive CORM-2 (iCORM-2), 1.6 mmol/L iCORM-2, with six samples in each group. After being cultured for 0, 3, 5, 8, 10, 12, 16, 20, 24, 27, 30, 48 hours, proliferative vitality of E. coli was examined (denoted as absorption value under 600 nm wavelength), and bacteria number was counted. Other standard strains of E. coli ATCC 25922 were divided into groups F (without addition) and G (cultured with 0.8 mmol/L CORM-2), the expressions of genes fliA, dnaK, marA, and waaQ related to E. coli were detected by quantitative real-time (qRT)-PCR. (2) In vivo experiments. Other standard strains of E. coli ATCC 25922 were grouped as A', B', C', D', and E' and treated with the same method as that in groups A, B, C, D, and E, and 0.5 mL bacterial liquid of each group were collected when the absorption value of bacterial liquid in group A' was equal to 0.4 (under 600 nm wavelength). Seventy-two C57BL/6 mice were divided into groups, namely blank control (without treatment), H, I, J, K, and L according to the random number table, with 12 mice in each group. The mice in the latter 5 groups were intraperitoneally injected with 0.5 mL bacterial suspension of groups A', B', C', D', and E' respectively. After injection, general condition of mice in groups H, I, J, K, and L was observed. The serum levels of TNF-α and IL-6 were determined at post injection hour (PIH) 6, 12. The liver and lung samples were harvested for determination of myeloperoxidase (MPO) activity at PIH 12. The same process was carried out in blank control group. Data were processed with repeated measure analysis of variance (ANOVA), factorial design ANOVA, one-way ANOVA, and t test.</p><p><b>RESULTS</b>(1) In vitro experiments. Compared with those of groups A and D, the proliferative vitality and bacteria number of E. coli in group B were all decreased (with F values respectively 1170.80, 217.52, P values all below 0.01). Compared with those of groups A and E, the proliferative vitality and bacteria number of E. coli in group C were also obviously decreased (with F values respectively 7948.34, 14 432.85, P values all below 0.01). Compared with those in group F, the expression of fliA was downregulated, while the expressions of dnaK, marA, and waaQ were upregulated in group G (with t values 30.28, -165.54, -168.88, -187.28, P values all below 0.01). (2) In vivo experiments. Symptoms including listlessness and tachypnea were observed in mice in groups H, K, and L, and they were ameliorated or not obvious in groups I and J. At PIH 6, the serum levels of TNF-α and IL-6 in groups H and K were respectively (647.3 ± 3.8) pg/mL, (3.44 ± 0.22) ng/mL and (639.3 ± 0.8) pg/mL, (2.47 ± 0.32) ng/mL, which were obviously higher than those in group I [(124.6 ± 10.7) pg/mL, (1.03 ± 0.16) ng/mL, with t values from 15.22 to 84.03, P values all below 0.01]. The serum levels of TNF-α and IL-6 in group J at PIH 6, 12 were also obviously decreased as compared with those in groups H and L (with t values from 19.27 to 245.34, P values all below 0.01). MPO activity of liver and lung tissues were significantly attenuated in group I at PIH 12 as compared with those in groups H and K, and it was also attenuated in group J when compared with those in groups H and L (with t values respectively from 17.36 to 18.92 and 2.35 to 3.61, P values all below 0.01).</p><p><b>CONCLUSIONS</b>CORM-2 can obviously inhibit the vitality and toxicity of E. coli, which might be attributable to regulation of expressions of genes fliA, dnaK, marA, and waaQ of E. coli.</p>

Inhibitory effect of extrinsic CO-releasing molecules-2 (CORM-2) on inflammatory responses in mice with LPS-induced acute lung injury / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of extrinsic CO-releasing molecules-2 (CORM-2) on attenuating pulmonary injury and the inflammatory response in LPS-induced acute lung injury of mice.</p><p><b>METHODS</b>Fifty-three mice were assigned to four groups. Mice in sham group (n= 8) underwent sham inhalation, whereas mice in ALI (n= 15) received inhalation of LPS for 30 min, mice in ALI+iCORM (n= 15) underwent LPS inhalation with immediate administration of inactive CORM-2 (8 mg/kg, i.v.), mice in ALI+CORM (n= 15) underwent LPS inhalation with immediate administration of CORM-2 (8 mg/kg, i.v.). PMN accumulation (MPO assay) in mice lungs and TNF-α and IL-1 β in BAL fluid were determined. Activation of NF-kB and expression level of ICAM-1 in the lung were assessed.</p><p><b>RESULT</b>Treatment of ALI mice with CORM-2 attenuated PMN accumulation and prevented activation of NF-kB in the lung. This was accompanied by a decrease of the expression of ICAM-1. In parallel, CORM-2 markedly decreased the production of inflammatory mediators in BAL fluid.</p><p><b>CONCLUSION</b>CORM-2 attenuates the inflammatory response in the lung of LPS-induced ALI by decreasing leukocyte sequestration and interfering with NF-kB activation, expression of ICAM-1 and therefore suppressing endothelial cells pro-adhesive phenotype.</p>

Atelocollagen-mediated small interfering RNA delivery for effective gene silencing in rat vein grafts / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of using small interfering RNA targeting TF as a therapy for vein graft failure.</p><p><b>METHODS</b>External jugular vein to carotid artery interposition vein grafts, which were applied to a low flow condition, were made in 120 Sprague-Dawley rats weighing 260 to 300 g. These rats were randomly divided into 4 groups, 30 rats each group. Group A was atelocollagen-TF Stealth Select RNAi group. Group B was atelocollagen-TF Stealth RNAi group. Group C was atelocollagen group. Group D was control group. Small interfering RNA mixed with atelocollagen was administrated to the external wall of grafted veins. The TF protein expression of vein grafts was analyzed by Western blot at 1, 3, 7, 14, and 28 d postoperatively, and by immunochemistry at 3 d postoperatively. The proliferation index was determined at 14 d postoperatively. Neointimal hyperplasia was evaluated at 28 d postoperatively. BLOCK-iT fluorescent oligo was used to confirm its stability and successful transfer into the vein graft wall at 3 and 7 d postoperatively for another group (n=12).</p><p><b>RESULTS</b>Fluorescence of BLOCK-iT fluorescent oligo could be detected in the graft wall even at 7 d postoperatively. Knockdown of the TF expression was achieved by perivascular application of siRNA using atelocollagen. Compared with control group, the intima thickness at 28 d after grafting was significantly reduced (P < 0.05). This phenomenon was preceded by significant reduction of cell proliferation in siRNA-treated grafts at 14 d postoperatively (P < 0.05).</p><p><b>CONCLUSION</b>The expression of TF in vein grafts can be effectively inhibited by specific siRNAs using a atelocollagen-based nonviral delivery approach in vivo, so that the neointimal thickening can be prevented. Transplants;</p>

Simultaneous off-pump coronary artery bypass surgery and pulmonary resection / 中华外科杂志

<p><b>OBJECTIVE</b>To summarize the experience of combined off-pump coronary artery bypass grafting (OPCAB) and pulmonary resection.</p><p><b>METHODS</b>Seven patients with unstable angina or a history of myocardial infarction and pulmonary disease underwent combined OPCAB and pulmonary resection. All of them underwent coronary angiography, and neither coronary angioplasty nor stenting was feasible. OPCAB preceded the lung resections. The preferred approach to the heart and lung was by sternotomy. Left upper lobectomy was performed in 2 patients, right upper lobectomy was performed in 1 patient, right lower lobectomy was performed in 1 patient, right upper and middle bilobectomy was performed in 1 patient, left lung volume reduction surgery (LVRS) was performed in 1 patient and bilateral LVRS was performed in 1 patient.</p><p><b>RESULTS</b>There were no hospital mortality in this group of patients, however there were one late death. Sternal dehiscence occurred in 1 patient which was observed with a need for re-sternotomy and atrial fibrillation was observed in 1 patient. Five patients were diagnosed as malignant tumor by pathology test, and 2 patients were severe chronic obstructive pulmonary disease (COPD). Follow-up ranging from 2 months to 31 months was available for these patients. None of the patients showed evidence of myocardial ischemia after surgery. In one patient, who underwent right upper and middle bilobectomy, local recurrence was found at 19 months after surgery.</p><p><b>CONCLUSIONS</b>OPCAB carried out simultaneously with lung resection is a safe and effective approach in patients diagnosed with concomitant coronary artery and pulmonary disease. OPCAB may decrease the incidence of postoperative complications.</p>