Objective To analyze the expression and detection significance of community acquired respiratory distress syndrome(CARDS)toxin in bronchoalveolar lavage fluid(BALF)in children with severe Mycoplasma pneumoniae pneumonia(SMPP).Methods The BALF samples of children diagnosed with Mycoplasma pneumoniae pneumonia(MPP)were collected from the Department of Pulmonology,Children's Hospital Affiliated to Capital Institute of Pediatrics from August 2021 to April 2022,the relative expression of CARDS toxins was determined by quantitative realtime polymerase chain reaction(qPCR).The patients were divided into SMPP group and non-SMPP group(control group).The relative expression of CARDS toxin and other clinical indicators were compared between the two groups.The risk factors of SMPP were analyzed.The predictive value of the indicators was analyzed using receiver operating characteristic(ROC)curves.Results ①The relative expression level of CARDS toxin(Z=-6.151),D-dimer(Z=-5.102)and lactate dehydrogenase(LDH)(Z=-6.337)in SMPP group were significantly higher than those in control group(P<0.001).The white blood cell(Z=-2.155),neutrophil ratio(t=2.988)and C-reactive protein(CRP)(Z=-3.360)were higher than those in control group(P<0.05).②The relative expression of CARDS toxin was positively correlated with serum LDH(r=0.258,P<0.05).③Binary Logistic regression analysis showed that fever time(OR=1.271,95% CI:1.079-1.497),LDH(OR=1.011,95% CI:1.005-1.017)and CARDS toxin(OR=6.210,95% CI:2.646-14.577)were risk factors for SMPP.Conclusion CARDS toxin was closely related to MPP disease,and was significantly increased in SMPP group.CARDS toxin was one of the independent risk factors for SMPP and positively correlated with serum LDH level.It can be used as a good indicator to evaluate the condition,combined with fever time and LDH has clinical significance.

Objective To analyze the expression and detection significance of community acquired respiratory distress syndrome(CARDS)toxin in bronchoalveolar lavage fluid(BALF)in children with severe Mycoplasma pneumoniae pneumonia(SMPP).Methods The BALF samples of children diagnosed with Mycoplasma pneumoniae pneumonia(MPP)were collected from the Department of Pulmonology,Children's Hospital Affiliated to Capital Institute of Pediatrics from August 2021 to April 2022,the relative expression of CARDS toxins was determined by quantitative realtime polymerase chain reaction(qPCR).The patients were divided into SMPP group and non-SMPP group(control group).The relative expression of CARDS toxin and other clinical indicators were compared between the two groups.The risk factors of SMPP were analyzed.The predictive value of the indicators was analyzed using receiver operating characteristic(ROC)curves.Results ①The relative expression level of CARDS toxin(Z=-6.151),D-dimer(Z=-5.102)and lactate dehydrogenase(LDH)(Z=-6.337)in SMPP group were significantly higher than those in control group(P<0.001).The white blood cell(Z=-2.155),neutrophil ratio(t=2.988)and C-reactive protein(CRP)(Z=-3.360)were higher than those in control group(P<0.05).②The relative expression of CARDS toxin was positively correlated with serum LDH(r=0.258,P<0.05).③Binary Logistic regression analysis showed that fever time(OR=1.271,95% CI:1.079-1.497),LDH(OR=1.011,95% CI:1.005-1.017)and CARDS toxin(OR=6.210,95% CI:2.646-14.577)were risk factors for SMPP.Conclusion CARDS toxin was closely related to MPP disease,and was significantly increased in SMPP group.CARDS toxin was one of the independent risk factors for SMPP and positively correlated with serum LDH level.It can be used as a good indicator to evaluate the condition,combined with fever time and LDH has clinical significance.

Objective:To investigate the value of community acquired respiratory distress syndrome (CARDS) toxin combined with IFN-γ in the early prediction of refractory Mycoplasma pneumoniae pneumonia (RMPP). Methods:A total of 100 children diagnosed with Mycoplasma pneumoniae pneumonia (MPP) were selected as the study objects. Among them, 36 cases were diagnosed with RMPP and 64 cases without RMPP were set as control group. The general data, clinical symptoms, signs, laboratory tests and internal and external pulmonary complications were compared between the two groups. The independent risk factors for RMPP was analyzed by Binary Logistic regression, and receiver operating characteristic (ROC) curve was used to analyze the predictive value of the indicators. Results:The fever time and hospital stay in RMPP group were significantly longer than those in control group, the relative expression of CARDS toxin, neutrophil ratio, C-reactive protein, D-dimer, lactate dehydrogenase, IL-6 and IFN-γ levels in RMPP group were higher than those in control group, and the differences were statistically significant. Binary Logistic regression analysis showed that long fever duration, increased expression of IFN-γ and CARDS toxin and extrapulmonary complications were independent risk factors for RMPP. ROC curves were drawn for the three continuous variables of fever time, IFN-γ, and relative expression of CARDS toxin, the sensitivity and specificity of the combined prediction of the three indexes were 86.1% and 73.4%, respectively.Conclusions:CARDS toxin is closely related to MPP. Fever duration, IFN-γ, relative expression of CARDS toxin and extrapulmonary complications are independent risk factors for RMPP. Especially when the fever time is more than 10.5 d, IFN-γ is more than 31.79 pg/ml, and the relative expression of CARDS toxin is more than 1.18, the possibility of RMPP should be noted.

Objective:To investigate the value of community acquired respiratory distress syndrome (CARDS) toxin combined with IFN-γ in the early prediction of refractory Mycoplasma pneumoniae pneumonia (RMPP). Methods:A total of 100 children diagnosed with Mycoplasma pneumoniae pneumonia (MPP) were selected as the study objects. Among them, 36 cases were diagnosed with RMPP and 64 cases without RMPP were set as control group. The general data, clinical symptoms, signs, laboratory tests and internal and external pulmonary complications were compared between the two groups. The independent risk factors for RMPP was analyzed by Binary Logistic regression, and receiver operating characteristic (ROC) curve was used to analyze the predictive value of the indicators. Results:The fever time and hospital stay in RMPP group were significantly longer than those in control group, the relative expression of CARDS toxin, neutrophil ratio, C-reactive protein, D-dimer, lactate dehydrogenase, IL-6 and IFN-γ levels in RMPP group were higher than those in control group, and the differences were statistically significant. Binary Logistic regression analysis showed that long fever duration, increased expression of IFN-γ and CARDS toxin and extrapulmonary complications were independent risk factors for RMPP. ROC curves were drawn for the three continuous variables of fever time, IFN-γ, and relative expression of CARDS toxin, the sensitivity and specificity of the combined prediction of the three indexes were 86.1% and 73.4%, respectively.Conclusions:CARDS toxin is closely related to MPP. Fever duration, IFN-γ, relative expression of CARDS toxin and extrapulmonary complications are independent risk factors for RMPP. Especially when the fever time is more than 10.5 d, IFN-γ is more than 31.79 pg/ml, and the relative expression of CARDS toxin is more than 1.18, the possibility of RMPP should be noted.

The clinical and genetic data of a child with cystic fibrosis who received double lung transplantation were retrospectively analyzed and related problems of cystic fibrosis lung transplantation were discussed.The child was diagnosed with cystic fibrosis in the Department of Respiratory, the Affiliated Children′s Hospital of Capital Institute of Pediatrics at the age of 6 years for the first time.The patient was hospitalized repeatedly for exacerbation of pulmonary infection, with no significant clinical improvement and progressive aggravation of malnutrition.In October 2019, the patient received double lung transplantation at the age of 17 years and 1 month.Two years after surgery, the clinical symptoms and quality of life have been notably improved, and no transplant-related complications were observed.Lung transplantation is an effective treatment for children with advanced cystic fibrosis lung disease.

Objective To explore the clinical features of children infected with macrolide-resistant (MR) Mycoplasma pneumonia(MP) isolates and genetic typing of all isolates.Methods Polymerase chain reaction(PCR) of MP positive in 96 nasopharyngeal or bronchoalveolar lavage fluid (BALF) samples were collected from patients diagnosed as MP pneumonia in the Affiliated Children's Hospital of the Capital Institute of Pediatrics from January 2013 to October 2015.Fifty-five cases were male,41 cases were female;19 cases (19.8％) were 1 to 3 years old,18 cases (18.7％) were more than 3 to 5 years old,59 cases(61.5％) were more than 5 to 13 years and 2 months old.These samples were tested for MR associated mutations in the 23S rRNA of MP,and were divided into the MR group and the macrolide-sensitive (MS) group.Furthermore,the genotype of all the isolates were performed by conducting P1-restriction fragment length polymorphism(P1-RFLP) analysis and multiple-locus variable-number tandem-repeat analysis (MLVA) method.The clinical characteristics including the age,gender,hospitalization duration,symptoms,signs,fever duration,fever duration after macrolide therapy,white blood cell count,C-reactive protein (CRP),chest X-ray and/or chest computed tomography,which were compared between different groups.SPSS 11.5 software was used to analyze the statistical data.Statistical significance was determined at the 0.05 level of a two-tailed test.Results MR mutations were identified in the 23S rRNA gene in 81 specimens (84％),and the 96 specimens were divided into MR group(81 cases) and MS group (15 cases).There were statistical differences in fever duration,hospitalization duration,the incidence of complications and CRP level between the MR group and MS group (t =2.061,Z =-3.368,x2 =5.856,Z =-2.165,all P ＜ 0.05).There were no statistical differences in age,white blood cell count,consolidation percentage on chest radiography and fever duration after macrolide therapy(all P ＞ 0.05).All the 96 isolates were performed by adopting P1-RFLP typing,but 5 isolates were not typed successfully,while 81 cases (89.0％) isolates were typed as P1-Ⅰ and 10 isolates(11％) were typed as P1-Ⅱ c.The hospitalization duration and the fever duration after macrolide therapy in the P1-Ⅰ were longer than the P1-Ⅱ c group,and the difference was statistically significant (Z =-2.197,2.237,all P ＜ 0.05).There were no statistical differences in age,fever duration,white blood cell count,CRP level,consolidation percentage on chest radiography and the incidence of complications (all P ＞ 0.05).Seventy-three cases (90％) of P1-Ⅰ group were MR isolates,8 cases (10％) were MS isolates;3 cases (30％) of the P1-]Ⅱ c group were MR isolates,7 cases (70％) were MS isolates.The MR isolates in P1-Ⅰ group were much more than P1-Ⅱ c group.There was obvious statistical difference in the proportion of MR isolates between 2 groups (x2 =19.209,P ＜ 0.01).All the 96 isolates were performed by modified MLVA typing,82 cases(85.5％) were typed as M4-5-7-2,11 cases(11.5％) were typed as M3-5-6-2,1 case (1.0％) was typed as M4-5-5-2,and 2 cases (2.0％) were typed as M4-5-6-2.Because there were less cases of the M4-5-5-2 and M4-5-6-2 type,only the clinical data of M4-5-7-2 and M3-5-6-2 group were compared.The hospitalization duration and the fever duration after macrolide therapy in the M4-5-7-2 group were longer than the M3-5-6-2 group,and the difference was statistically significant(Z =-2.406,-4.472,all P ＜ 0.05).There were no statistical differences in age,fever duration,white blood cell count,CRP level,consolidation percentage on chest radiography and the incidence of complications(all P ＞ 0.05).Seventy-four cases (90％) of the M4-5-7-2 group were MR isolates,8 cases (10％) were MS isolates;4 cases (36％) of the M3-5-6-2 group were MR isolates,7 cases (64％) were MS isolates.The MR isolates in M4-5-7-2 group were much more than M3-5-6-2 group.There was obviously statistical difference in the proportion of MR isolates between 2 groups (x2 =17.022,P ＜ 0.01).Conclusions In the MR group,the children had longer fever duration and hospitalization duration,higher incidence of complications and higher CRP level than those in the MS group.The MR rates of MP in China was high.P1-Ⅰ and M4-5-7-2 are the predominate genotypes.There may be a correlation between genotype and MR.

ABSTRACT:Objective To explore the protective effect of the antioxidant N‐acetylcysteine (NAC) on the retinal nerve tissue of early diabetic rats .Methods We randomly divided 60 healthy adult Sprague‐Dawley (SD) rats weighing between 180 g and 220 g into 2 groups:normal control (CON , n=20) and diabetic (DM , n=40) .By intraperitoneal injection of streptozotocin (60 mg/kg) ,the model of diabetic rats was established .The rats were considered diabetic only when they had hyperglycemia (set at ≥16 .7 mmol/L) (32) .The CON group was injected with the same amount of citric acid and sodium citrate buffer solution .After successful model establishment ,the diabetic rats were randomly divided into 1‐month diabetes group and 2‐month diabetes group ,with 16 rats in each group .The left eye of each experimental diabetic rat was set for diabetes control group (D) while the right eye was set as NAC treatment group (NAC) .At 2 weeks of diabetes ,4μL (1 .6μg/μL) of NAC was injected into the vitreous chamber of NAC group and 4μL (0 .01 mmol/L) of PBS was injected into the vitreous chamber of the other diabetic rats .The thickness changes of outer nuclear layer retina was observed by HE ,ultrastructural changes of retinal ganglion cells were observed under the transmission electron microscope ,and the number of retinal ganglion cells was detected by immunofluorescence method .Results At different time points ,retina outer nuclear layer in NAC group was thicker than in D group (P<0 .01) .However ,the NAC group and the CON group did not differ (P>0 .05) .Under the transmission electron microscope ,NAC group had more retinal ganglion cell organelles ,higher electron density of the cytoplasm ,and milder mitochondria swelling than D group .The NAC group did not differ from CON group in the ultrastructure of retinal ganglion cells . NAC group had an increased number of retinal ganglion cells at different time points compared with the D group (P<0 .01) ,but the NAC and CON groups did not differ in the number of retinal ganglion cells (P> 0 .05) .Conclusion The antioxidant N‐acetylcysteine has a protective effect on the retinal nerve tissue of early diabetic rats .

The newly emerged Middle East respiratory syndrome coronavirus (MERS-CoV) is a highly pathogenic respiratory virus with pathogenic mechanisms that may be driven by innate immune pathways. The goal of this study is to characterize the expression of the structural (S, E, M, N) and accessory (ORF 3, ORF 4a, ORF 4b, ORF 5) proteins of MERS-CoV and to determine whether any of these proteins acts as an interferon antagonist. Individual structural and accessory protein-coding plasmids with an N-terminal HA tag were constructed and transiently transfected into cells, and their native expression and subcellular localization were assessed using Wes tern blotting and indirect immunofluorescence. While ORF 4b demonstrated majorly nuclear localization, all of the other proteins demonstrated cytoplasmic localization. In addition, for the first time, our experiments revealed that the M, ORF 4a, ORF 4b, and ORF 5 proteins are potent interferon antagonists. Further examination revealed that the ORF 4a protein of MERS-CoV has the most potential to counteract the antiviral effects of IFN via the inhibition of both the interferon production (IFN-β promoter activity, IRF-3/7 and NF-κB activation) and ISRE promoter element signaling pathways. Together, our results provide new insights into the function and pathogenic role of the structural and accessory proteins of MERS-CoV.
Cell Line ; Coronavirus ; genetics ; pathogenicity ; Genes, Viral ; Humans ; Interferons ; antagonists & inhibitors ; Open Reading Frames ; Recombinant Proteins ; genetics ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism ; Viral Regulatory and Accessory Proteins ; genetics ; metabolism ; Viral Structural Proteins ; genetics ; metabolism

ObjectiveTo investigate the effects of grape seed proanthocyanidin extracts (GSPE) on advanced glycation end product receptor (RAGE),NF-Κb and connective tissue growth factor (CTGF) in the myoeardium of diabetic rats.MethodsTotal 30 streptozotocin (STZ) induced diabetic rats were randomly divided into 2 groups:diabetic group (DM1,n-15) and GSPE (250 mg/ kg,i.g) treated diabetic group (DM2,n= 15).Another two control groups:normal rats(C1,n= 10)and normal rats treated with GSPE (250 mg/kg,i.g) (C2 group,n= 10) were also observed.After 24 weeks,blood was collected to measure fasting plasma glucose (FBG) and RAGE.The protein expression of NF-Κb was determined in myocardial tissue by immunohistochemical staining and Western blot.The protein expressions of RAGE and CTGF were measured by Western blot.Results The levels of FBG and RAGE were significantly higher in diabetic rats than in control rats (P＜0.05).After GSPE treatment,RAGE level significantly reduced (P＜0.05),but FBG had no change in diabetic rats.The protein expressions of RAGE,NF-Κb and CTGF in the myocardial tissue of diabetic rats had marked increase compared with control rats (P＜ 0.05),however,their levels significantly reduced after GSPE treatment (P＜0.05).ConclusionsGSPE may protect diabetic rats against cardiomyopathy,possibly by decreasing the protein expressions of RAGE,NF-Κb and CTGF.

potential use in clinical therapy and prognosis estimation of the tumor.