Relationship between intestinal microbiota heterogeneity and acquired myasthenia in septic mice / 中华麻醉学杂志

Objective:To evaluate the relationship between intestinal microbiota heterogeneity and acquired myasthenia in septic mice.Methods:Eighty SPF healthy male C57BL/6 mice, weighing 18-20 g, aged 6-8 weeks, were included. Forty mice were selected to prepare a sepsis model by intraperitoneal injection of lipopolysaccharide (LPS) 10 mg/kg. Sepsis-sensitive mice (state of dying or even death within 24 h after developing the model) and sepsis-resistant mice (survival for 7 days and recovery) were screened. The feces from donor mice were collected to make fecal bacteria fluid. Forty mice were selected and divided into 4 groups ( n=10 each) by the random number table method: control group (C group), antibiotic group (ABX group), resistant group (Res group), and sensitive group (Sen group). Group C received no treatment. In ABX group, compound antibiotics were given by intragastric gavage once a day for 5 consecutive days. In Res group, the fecal solution from sepsis-resistant mice 150 μl was given by gavage once a day for 3 consecutive days starting from 5 days after gavage administration of compound antibiotics, and then LPS 15 mg/kg was intraperitoneally injected. In Sen group, the fecal solution from sepsis-sensitive mice was given by gavage for 3 days (using the same method as previously described in Res group) starting from 5 days after gavage administration of compound antibiotics, and then LPS 15 mg/kg was intraperitoneally injected. The severity of sepsis was assessed and scored at 24 h after developing the model. The four limb grip strength was measured after fecal bacteria transplantation and at 24 h after developing the sepsis model. The Compound Muscle Action Potential (CMAP) of the gastrocnemius was measured at 24 h after developing the sepsis model. Then blood samples were collected for determination of the levels of interleukin-1 (IL-1), IL-6 and tumor necrosis factor α (TNF-α) in serum by enzyme-linked immunosorbent assay. The tissues of anterior tibial muscle and gastrocnemius muscle were obtained for determination of the expression of muscle-specific ring finger protein 1 (MuRF-1) and muscle atrophy box F protein (MAFbx) by Western blot. The diameter and cross-sectional area of gastrocnemius muscle fibers were measured after HE staining. The feces of mice were collected at 3 days after fecal bacteria transplantation, and DNA was extracted from mouse fecal samples for 16S rDNA sequencing and untargeted metabolomics analysis. Results:Compared with C group, the score for severity of sepsis was significantly increased, the four limb grip strength was decreased after developing the sepsis model, the serum IL-6 concentration was increased, and the diameter and cross-sectional area of gastnemius muscle fibers were decreased ( P<0.05), and no statistically significant changes were found in the other parameters in Res group, and no statistically significant changes were found in the indexes mentioned above in ABX group ( P>0.05). Compared with C group and Res group, the score for severity of sepsis was significantly increased, the four limb grip strength was decreased, the latency of CMAP was prolonged, and the amplitude of CMAP was decreased, the serum concentrations of IL-1, IL-6 and TNF-α were increased, the diameter and cross-sectional area of gastrocnemius muscle fibers were decreased, and the expression of MuRF-1 and MAFbx in anterior tibial muscle was up-regulated after developing the sepsis model in Sen group ( P<0.05). 16S rDNA sequencing of intestinal flora: Compared with Sen group, no significant change was found in Chao1 index, Good-coverage index and Simpson index in Res group ( P>0.05), Shannon index was increased ( P<0.05), and no significant change was found in β diversity in Res group ( P>0.05). LDA analysis showed that f_Akkermarisiaceae, o_Verrucomicrobiales, g_Akmansia, c_Verrucomicrobiae and p_Verrucomicrobiota were significantly enriched in Sen group, and g_Enterococcus and f_Enterococcaceae were significantly enriched in Res group ( P<0.05). Nontargeted metabolic analysis: The metabolite profiles in Res group and Sen group suggested that the model was well developed (R2Y>Q2Y). Compared with Sen group, 90 metabolites was significantly up-regulated and 88 down-regulated, significantly up-regulated metabolic substances including vitamin K1, gamma-tocopherol, and taurine in Res group ( P<0.05). The differential metabolite KEGG enrichment pathway included 2-oxocarboxylic acid metabolism and ubiquinone and other terpenoid-quinone biosynthesis ( P<0.05). Conclusions:Intestinal flora heterogeneity may be involved in the pathogenesis of acquired myasthenia in septic mice.

Enhanced endoplasmic reticulum RyR1 receptor phosphorylation leads to diaphragmatic dysfunction in septic rats / 南方医科大学学报

OBJECTIVE@#To explore the role of endoplasmic reticulum ryanodine receptor 1 (RyR1) expression and phosphorylation in sepsis- induced diaphragm dysfunction.@*METHODS@#Thirty SPF male SD rats were randomized equally into 5 groups, including a sham-operated group, 3 sepsis model groups observed at 6, 12, or 24 h following cecal ligation and perforation (CLP; CLP-6h, CLP-12h, and CLP-24h groups, respectively), and a CLP-24h group with a single intraperitoneal injection of KN- 93 immediately after the operation (CLP-24h+KN-93 group). At the indicated time points, diaphragm samples were collected for measurement of compound muscle action potential (CMAP), fatigue index of the isolated diaphragm and fitted frequencycontraction curves. The protein expression levels of CaMK Ⅱ, RyR1 and P-RyR1 in the diaphragm were detected using Western blotting.@*RESULTS@#In the rat models of sepsis, the amplitude of diaphragm CMAP decreased and its duration increased with time following CLP, and the changes were the most obvious at 24 h and significantly attenuated by KN-93 treatment (P < 0.05). The diaphragm fatigue index increased progressively following CLP (P < 0.05) irrespective of KN- 93 treatment (P>0.05). The frequency-contraction curve of the diaphragm muscle decreased progressively following CLP, and was significantly lower in CLP-24 h group than in CLP-24 h+KN-93 group (P < 0.05). Compared with that in the sham-operated group, RyR1 expression level in the diaphragm was significantly lowered at 24 h (P < 0.05) but not at 6 or 12 following CLP, irrespective of KN-93 treatment; The expression level of P-RyR1 increased gradually with time after CLP, and was significantly lowered by KN-93 treatment at 24 h following CLP (P < 0.05). The expression level of CaMKⅡ increased significantly at 24 h following CLP, and was obviously lowered by KN-93 treatment (P < 0.05).@*CONCLUSION@#Sepsis causes diaphragmatic dysfunction by enhancing CaMK Ⅱ expression and RyR1 receptor phosphorylation in the endoplasmic reticulum of the diaphragm.

Effect of alprostadil in the prevention of contrast nephropathy in elderly patients with coronary heart disease after interventional treatment / 中国基层医药

Objective To investigate the effect of alprostadil in the treatment of interventional angiography in elderly patients with coronary heart disease after contrast nephropathy prevention.Methods From February 1,2014 to January 31,2017,60 elderly patients with coronary heart disease with interventional therapy in Yuncheng Central Hospital of Shanxi Province were divided into two groups,with 30 cases in each group.The control group received hydration therapy,the observation group was given alprostadil combined with hydration therapy.The biochemical index changes before surgery and three days after surgery and incidence of contrast nephropathy were compared between the two groups.Results Before surgery and three days after surgery,the tumor necrosis factor alpha,beta 2 microglobulin, urea and creatinine in the two groups were not changed significantly.Three days after surgery,24h urine protein, superoxide dismutase,glutathione peroxidase,interleukin －6 and C reactive protein levels in the two groups were increased,the increase amplitude of the observation group was smaller.Three days after surgery,the five indicators of the observation group were (185.54 ±86.47)mg,(2.01±1.32)mg/L,(6.18 ±2.13)g/L,(135.56 ±41.58)ng/L, (1.21±1.05 )mg/L,respectively,the differences were ststistically significant between the two groups (t ＝1.21, 1.24,1.50,1.26,1.22;P＝0.03,0.03,0.04,0.03,0.04).Three days after surgery,the creatinine clearance rate of the observation group was (98.67 ±21.56)mL/min,which was higher than that of the control group(t＝2.71,P＝0.01).The incidence rate of contrast induced nephropathy in the observation group (3.33%)was lower than that in the control group(χ2＝5.19,P＝0.02).Conclusion Alprostadil can effectively prevent contrast nephropathy in elderly patients with coronary heart disease after interventional therapy.

The effect of oral care on the prevention of ventilator associated pneumonia in patients with severe traumatic brain injury by aspiration of subglottic secretion combined with 2%acetic acid chloride solution / 中国实用护理杂志

Objective To explore the effect of oral care on the prevention of ventilator associated pneumonia in patients with severe traumatic brain injury by aspiration of subglottic secretion combined with 2% acetic acid chloride solution. Methods A total of 100 cases of ICU patients with mechanical ventilation for longer than 48 hours were divided into group B and group C using a random number table method according to tracheotomy mechanical ventilation time sequence number from January 2014 to June 2015. Another 50 ICU patients in group A (as the case for retrospective analysis, from April to November , 2013) were treated by routine tracheal suction, normal saline cotton for oral care, group B were treated with aspiration of subglottic secretion, normal saline cotton for oral care, group C were treated with aspiration of subglottic secretion combined with 2% acetic acid chloride solution for oral cleaning. And then VAP occurrence rate, plaque index, plaque clearance were evaluated. Results The incidence rates of early onset VAP in three groups were 14% (7/50), 8% (4/50), 4% (2/50), and incidence rates of late onset VAP were 22% (11/50),12% (6/50), 8% (4/50) respectively, the differences were significant (χ2=3.65, 3.88,P<0.01). The plaque index and plaque clearance rates of the three groups on the third day were 2.14± 0.35, (40.12 ± 13.11)%, 2.10 ± 0.33, (39.17 ± 14.21)%, 1.03 ± 0.24, (60.12 ± 15.07 )%(F=12.21,χ2=3.82,P<0.01);while on the 7th day, they were 1.76 ± 0.47, (41.12 ± 13.34)%, 1.80 ± 0.32, (39.37 ± 12.21)%, 0.99 ± 0.15, (60.33 ± 14.25)%(F=13.14,χ2=4.20,P<0.01). Conclusions The incidence rate of VAP can be decreased by the combination of aspiration of subglottic secretion and oral cleaning with 2%acetic acid chloride solution.

The affect of body position on 32 biochemical indexes / 中华检验医学杂志

Objective To investigate the affect of body positions on biochemical indexes. Methods By autogenous contrast and cross matched survey, 107 volunteers divided into 3 season patches of winter, spring and summer, blood samples were drawn from the same part in both standing and lying positions。From19 persons, blood samples were collected respectively after standing and sitting for 15 min, lying for 15 min and 30 min and then sitting for another 15 min。 The blood samples were analyzed for 32 biochemical indexes on analyzer。Results 25 biochemical indexes in sitting position were significantly different from those in lying position (P0。05)。Conclusions Changing body position can result in obvious physiological variation of 28 biochemical indexes, particularly of those related to protein. Such result may lead to abnormality in some marginal values. It suggests body position should not be neglected in analyzing laboratory data.