Objective:To explore the effects and possible mechanism of intestinal microbiota on lung injury in mice with acute necrotizing pancreatitis (ANP).Methods:The experimental mice were randomly assigned to normal control group (CON group), ANP model group (ANP group) and intestinal germ-free group (ABX group), with 6 mice in each group. The ANP mouse model was constructed by intraperitoneal injection of caerulein (100 μg/kg, for 10 times) at an interval of 1 hour each time, followed by 10 mg/kg lipopolysaccharide injection. Mice in ABX group were treated by Abx solution (0.5 g/L vancomycin, 1 g/L neomycin, 1 g/L metronidazole, and 1 g/L ampicillin), 1 ml/100 g gavage for 28 days before preparation of the ANP model. The CON group was injected intraperitoneally with an equal volume of PBS. Histopathologic examination of the pancreas, lungs, and terminal ileum was routinely performed. Serum amylase levels were measured using enzymatic kinetic chemistry, and serum diamine oxidase (DAO) and lung tissue myeloperoxidase (MPO) activities were measured using ELISA assay. Expression of inflammatory factors, pyroptosis-related molecules in lung tissue and intestinal epithelial tight junction proteins was detected by fluorescence quantitative PCR. Western blotting was used to detect the expression of the pyroptosis molecules caspase-1 and GSDMD in lung tissue, and intestinal epithelial tight junction proteins. Changes of bacterial distribution in lung tissue were measured by fluorescence in situ hybridization. Results:The pathological scores of pancreatic tissue of CON, ANP, and ABX group were (0.67±0.26), (7.33±0.82), and (5.67±0.81); the pathological scores of lung tissue were (1.67±0.41), (5.67±0.41), and (3.58±0.58); the pathological scores of ileal tissue were (0.58±0.52), (3.83±0.75), and (4.33±0.82); the serum amylase levels were (403.95±93.11), (1037.24±126.77), and (647.32±145.90)U/L; the MPO levels in lung tissue were (0.23±0.03), (0.63±0.09), and (0.48±0.05)U/g. ABX group had significantly lower scores in pancreatic and lung tissues, serum amylase levels, and MPO levels in lung tissue compared to ANP group, and all the differences were statistically significant (all P value <0.05). The expression level in pancreatis tissue from CON, ANP and ABX group of IL-1β mRNA was 1.84±0.90, 36.26±5.56 and 16.65±6.43, IL-6 mRNA was 1.07±0.15, 2.90±0.42 and 1.34±0.62, TNF-α mRNA was 0.47±0.11, 0.76±0.11 and 0.46±0.07, HMGB1 mRNA was 0.38±0.02, 0.72±0.22 and 0.44±0.08, caspase-1 mRNA was 1.07±0.18, 2.04±0.31 and 0.85±0.54, ASC mRNA was 1.24±0.19, 5.68±0.41 and 3.89±1.47, GSDMD mRNA was 0.79±0.17, 0.94±0.14 and 0.61±0.08, IL-18 mRNA was 0.83±0.27, 4.17±0.79 and 3.57±0.03, respectively. The expression of IL-1β, IL-6, TNF-α, HMGB1, caspase-1, ASC, and IL-18 mRNA in lung tissue was significantly increased in ANP group compared to the CON group; conversely, ABX group showed a significant decrease in the expression of these markers compared to ANP group; and all the differences were statistically significant (all P values <0.05). The protein level of caspase-1 in lung tissue of CON, ANP and ABX group was 1.59±0.51, 2.28±0.13, 1.38±0.47, and that of GSDMD was 1.90±0.09, 2.20±0.07 and 1.76±0.27, respectively, which in ANP group were significantly higher than in CON group, but in ABX group was significantly lower than in ANP group, and all the differences were statistically significant (all P values <0.05). The serum DAO levels of CON, ANP, and ABX group were (0.06±0.15), (0.52±0.11) and (0.58±0.11) ng/ml; the expression level of ileum tissue of claudin1 mRNA and protein was 0.98±0.26, 0.42±0.18, 0.32±0.24 and 1.05±0.08, 0.82±0.09, 0.19±0.04; occludin mRNA and protein was 0.91±0.07, 0.31±0.05, 0.32±0.14 and 1.03±0.07, 0.61±0.04, 0.64±0.11; ZO-1 mRNA and protein was 1.01±0.08, 0.80±0.28, 0.60±0.28, and 0.86±0.10, 0.99±0.30, 0.62±0.30. The serum DAO level was significantly elevated in both ANP and ABX groups compared to the CON group. The mRNA and protein expression of claudin-1 and occludin in both ANP and ABX groups were significantly lower than those in CON group; the expression of claudin-1 in ABX group was significantly downregulated compared to ANP group; and all the differences were statistically significant (all P values <0.05). The relative fluorescence intensities of lung tissue in CON, ANP, and ABX groups were 0.03±0.01, 0.06±0.01, and 0.04±0.01, respectively, which in ANP group was significantly higher compared to CON group, but in ABX group was significantly lower than ANP group; all the differences were statistically significant (all P values <0.05). Conclusions:Intestinal microbiota may attenuate acute pancreatitis-associated acute lung injury by inhibiting the pyroptosis pathway in lung tissue.

Objective:To explore the effects and possible mechanism of intestinal microbiota on lung injury in mice with acute necrotizing pancreatitis (ANP).Methods:The experimental mice were randomly assigned to normal control group (CON group), ANP model group (ANP group) and intestinal germ-free group (ABX group), with 6 mice in each group. The ANP mouse model was constructed by intraperitoneal injection of caerulein (100 μg/kg, for 10 times) at an interval of 1 hour each time, followed by 10 mg/kg lipopolysaccharide injection. Mice in ABX group were treated by Abx solution (0.5 g/L vancomycin, 1 g/L neomycin, 1 g/L metronidazole, and 1 g/L ampicillin), 1 ml/100 g gavage for 28 days before preparation of the ANP model. The CON group was injected intraperitoneally with an equal volume of PBS. Histopathologic examination of the pancreas, lungs, and terminal ileum was routinely performed. Serum amylase levels were measured using enzymatic kinetic chemistry, and serum diamine oxidase (DAO) and lung tissue myeloperoxidase (MPO) activities were measured using ELISA assay. Expression of inflammatory factors, pyroptosis-related molecules in lung tissue and intestinal epithelial tight junction proteins was detected by fluorescence quantitative PCR. Western blotting was used to detect the expression of the pyroptosis molecules caspase-1 and GSDMD in lung tissue, and intestinal epithelial tight junction proteins. Changes of bacterial distribution in lung tissue were measured by fluorescence in situ hybridization. Results:The pathological scores of pancreatic tissue of CON, ANP, and ABX group were (0.67±0.26), (7.33±0.82), and (5.67±0.81); the pathological scores of lung tissue were (1.67±0.41), (5.67±0.41), and (3.58±0.58); the pathological scores of ileal tissue were (0.58±0.52), (3.83±0.75), and (4.33±0.82); the serum amylase levels were (403.95±93.11), (1037.24±126.77), and (647.32±145.90)U/L; the MPO levels in lung tissue were (0.23±0.03), (0.63±0.09), and (0.48±0.05)U/g. ABX group had significantly lower scores in pancreatic and lung tissues, serum amylase levels, and MPO levels in lung tissue compared to ANP group, and all the differences were statistically significant (all P value <0.05). The expression level in pancreatis tissue from CON, ANP and ABX group of IL-1β mRNA was 1.84±0.90, 36.26±5.56 and 16.65±6.43, IL-6 mRNA was 1.07±0.15, 2.90±0.42 and 1.34±0.62, TNF-α mRNA was 0.47±0.11, 0.76±0.11 and 0.46±0.07, HMGB1 mRNA was 0.38±0.02, 0.72±0.22 and 0.44±0.08, caspase-1 mRNA was 1.07±0.18, 2.04±0.31 and 0.85±0.54, ASC mRNA was 1.24±0.19, 5.68±0.41 and 3.89±1.47, GSDMD mRNA was 0.79±0.17, 0.94±0.14 and 0.61±0.08, IL-18 mRNA was 0.83±0.27, 4.17±0.79 and 3.57±0.03, respectively. The expression of IL-1β, IL-6, TNF-α, HMGB1, caspase-1, ASC, and IL-18 mRNA in lung tissue was significantly increased in ANP group compared to the CON group; conversely, ABX group showed a significant decrease in the expression of these markers compared to ANP group; and all the differences were statistically significant (all P values <0.05). The protein level of caspase-1 in lung tissue of CON, ANP and ABX group was 1.59±0.51, 2.28±0.13, 1.38±0.47, and that of GSDMD was 1.90±0.09, 2.20±0.07 and 1.76±0.27, respectively, which in ANP group were significantly higher than in CON group, but in ABX group was significantly lower than in ANP group, and all the differences were statistically significant (all P values <0.05). The serum DAO levels of CON, ANP, and ABX group were (0.06±0.15), (0.52±0.11) and (0.58±0.11) ng/ml; the expression level of ileum tissue of claudin1 mRNA and protein was 0.98±0.26, 0.42±0.18, 0.32±0.24 and 1.05±0.08, 0.82±0.09, 0.19±0.04; occludin mRNA and protein was 0.91±0.07, 0.31±0.05, 0.32±0.14 and 1.03±0.07, 0.61±0.04, 0.64±0.11; ZO-1 mRNA and protein was 1.01±0.08, 0.80±0.28, 0.60±0.28, and 0.86±0.10, 0.99±0.30, 0.62±0.30. The serum DAO level was significantly elevated in both ANP and ABX groups compared to the CON group. The mRNA and protein expression of claudin-1 and occludin in both ANP and ABX groups were significantly lower than those in CON group; the expression of claudin-1 in ABX group was significantly downregulated compared to ANP group; and all the differences were statistically significant (all P values <0.05). The relative fluorescence intensities of lung tissue in CON, ANP, and ABX groups were 0.03±0.01, 0.06±0.01, and 0.04±0.01, respectively, which in ANP group was significantly higher compared to CON group, but in ABX group was significantly lower than ANP group; all the differences were statistically significant (all P values <0.05). Conclusions:Intestinal microbiota may attenuate acute pancreatitis-associated acute lung injury by inhibiting the pyroptosis pathway in lung tissue.

Background:Acute lung injury(ALI)is the most common organ dysfunction in severe acute pancreatitis(SAP).Somatostatin analogue octreotide is a common used drug in acute pancreatitis.Aims:To explore the protective mechanism of octreotide on lung injury in SAP mice.Methods:In the first part,the experimental mice were randomly assigned into four groups.SAP model was induced by caerulin and lipopolysaccharide,and the mice were sacrificed 24 hours,48 hours and 72 hours after establishment.HE staining was used to observe the pathological score of pancreas and lung.Serum amylase and lung tissue myeloperoxidase(MPO)activity were detected.Real-time quantitative PCR was used to detect mRNA expressions of pyroptosis-related molecules apoptosis-associated speck-like protein containing a CARD(ASC),caspase-1,Gasdermin D(GSDMD),interleukin(IL)-1β,IL-18 and inflammatory factors IL-6,tumor necrosis factor(TNF)-α,high mobility group protein B1(HMGB1)in lung tissue.Western blotting was used to detect protein expressions of NOD-like receptor thermal protein domain associated protein 3(NLRP3),caspase-1,GSDMD and IL-1β in lung tissue.In the second part,mice were randomly divided into control group,SAP group,and octreotide group.HE staining was used to observe the pathological score of pancreas and lung.Serum amylase and lung tissue MPO activity were detected.Real-time quantitative PCR was used to detect mRNA expressions of pyroptosis-related molecules caspase-1,ASC,IL-1β,IL-18 and inflammatory factors IL-6,TNF-α,HMGB1.Immunofluorescence was used to detect protein expressions of NLRP3,caspase-1,GSDMD,ASC,IL-1β in lung tissue.Results:In the first part,compared with control group,pathological score of pancreas and lung tissue,serum amylase and MPO activity were significantly increased in SAP group(all P<0.05),mRNA expressions of pyroptosis-related molecules caspase-1,ASC,GSDMD,IL-1β,IL-18 and inflammatory factors IL-6,TNF-α,HMGB1 were significantly increased(all P<0.05),protein expressions of NLRP3,caspase-1,GSDMD and IL-1β in lung tissue were significantly increased(all P<0.05),especially in 24 hours after establishment group.In the second part,compared with SAP group,pathological score of pancreas and lung tissue,serum amylase were significantly decreased in octreotide group(all P<0.05),mRNA expressions of pyroptosis-related molecules caspase-1,ASC,IL-1β,IL-18 and inflammatory factors IL-6,TNF-α,HMGB1 were significantly decreased in lung tissue in octreotide group(all P<0.05),protein expressions of NLRP3,caspase-1,GSDMD,ASC and IL-1β in lung tissue were significantly decreased(all P<0.05).Conclusions:Cell pyroptosis is involved in the occurrence and development of lung injury in SAP mice,and octreotide may attenuate lung injury in SAP mice by inhibiting pyroptosis.

Objective:To understand the status quo and influencing factors of nurses' cognition on discharge preparation services, and to provide a basis for hospitals to further develop discharge preparation services and improve the quality of continuous nursing services.Methods:This study was a cross-sectional study. Clinical nurses from a ClassⅢ Grade A general hospital in Beijing were selected as research objects by the convenient sampling method in July 2020. A self-designed questionnaire (including knowledge, attitudes and behaviors related to discharge preparation services and influencing factors for implementation) was used to investigate. A total of 350 nurses participated in this survey, and the effective recovery rate of the questionnaire was 100%.Results:The mean score of discharge preparation service knowledge was (64.99±14.49) among 350 nurses. 42.9% of nurses said they had not heard of discharge preparation service, and 92.6% of nurses had not attended related training. 75.1% of the nurses thought it was necessary to carry out discharge preparation service, only 16.6% of the nurses had applied this model in clinical practice. There were statistically significant differences in the scores of nurses with different professional titles and knowledge of discharge preparation services ( P<0.05) . 46.3% of the nurses hoped to learn through online teaching. The main factors affecting discharge preparation service were heavy clinical nursing work and limited energy (86.9%) . Conclusions:Nurses have insufficient knowledge of discharge preparation service, so it is necessary to train nurses on discharge preparation service, so as to improve the quality of continuous nursing service and satisfaction of patients.