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Objective:To analyze the clinical characteristics and pathogenic distribution of severe pneumonia in adults in order to provide basis for clinical diagnosis and treatment.Methods:From June 2021 to April 2022, 145 patients with pneumonia admitted to the Department of Respiratory and Critical Care Medicine of the Second People's Hospital of Guangdong Province. According to whether they meet the diagnostic criteria for severe pneumonia, they were divided into severe ( n=63) and mild ( n=82) groups, and the clinical features between the two groups were compared. At the same time, the role of FilmArray detection in severe pneumonia was discussed. The measurement data were tested using independent sample t test or Mann-Whitney U test, and the counting data were tested using Chi-square test or Fisher exact probability method. Results:The age of the patients in the severe group was (72.67±1.71) years, male patients accounted for 84.1%, and the median hospitalization time was 16 days. Nine patients died in hospital; most of them had fever, shortness of breath, and change of consciousness, accompanied by hypertension, diabetes, cerebrovascular disease, chronic kidney disease, and tumor history. Compared with the mild group, the total number of leukocytes, neutrophil ratio, procalcitonin, and C-reactive protein were higher in the severe group, but the CD3 +, CD4 +, and CD8 + cell counts were lower ( P<0.05). The positive rate of FilmArray detection in the severe group was 81%, and the mixed infection of multiple bacteria accounted for 50%, which was higher than that of traditional culture ( P<0.05). The top four pathogens in severe group were Pseudomonas aeruginosa, Acinetobacter baumannii complex, Klebsiella pneumoniae, and Staphylococcus aureus, which were significantly higher than that in the mild group ( P<0.05). Resistance genes were detected in patients with severe disease, which was significantly higher than that in patients with mild disease (70.7% vs. 17.5%, P<0.05). Conclusions:Severe pneumonia is more common in elderly men, with more basic diseases and poor immunity. FilmArray has a high positive rate and can detect multiple pathogens, which may have a role in the rapid diagnosis of severe pneumonia.
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The β2m (Beta-2-microglobin) gene encodes a non-glycosylated protein that functions as an important component of major histocompatibility complexⅠ(MHCⅠ) for antigen presentation. To evade immune mediated clearance, human tumors and pathogens have adopted different strategies, including loss of MHCⅠexpression. Appropriate animal models are essential for understanding the mechanisms underpinning the clinical treatment of tumor and other human diseases. We constructed β2m knockout mice using CRISPR/Cas9 gene editing tool through embryo microinjection. Subsequently, genotyping and phenotyping of knockout mice were performed by PCR, qPCR, and flow cytometry. Mice genotyping showed that the coding region of the target gene was absent in the knockout mice. Real time PCR showed that mRNA level of β2m was significantly downregulated. Flow cytometry showed that the proportions of CD8+ killer T cells was significantly reduced in a variety of tissues and organs of the immune system. Taken together, we have successfully constructed a strain of β2m knockout mice, which will facilitate subsequent in vivo study on the function and mechanism of the β2m gene.
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Animals , Mice , Histocompatibility Antigens Class I , Mice, Inbred C57BL , Mice, Knockout , T-Lymphocytes, Cytotoxic , beta 2-Microglobulin/geneticsABSTRACT
Objective To explore the diagnostic value of virtual bronchoscopic navigation (VBN) for peripheral pulmonary lesions. Methods 200 cases with peripheral pulmonary lesions (0.8 cm < diameter < 4.0 cm) were divided into VBN and control group from June 2014 to June 2015. VBN group: VBN guided ultrafine bronchoscope to the target bronchus, control group: ultrafine bronchoscopy with chest CT as a reference guided to the target bronchus. Results There were no significant differences in the diagnosis rates between VBN group and control group in 200 patients (χ2 = 3.31, P = 0.069); But, the diagnosis rates with diameter more than 2.0 cm and less than or equal to 2.0 cm had statistically significant differences of VBN group and control group (χ2 = 13.45, 5.31, P = 0.000, 0.021, respectively); We also found that the mean time of biopsy tool reach to the lesions had significant differences between the two groups (P = 0.047). There were no significant differences in total checking time and complications (P = 0.230, 0.960, respectively). Conclusions Virtual bronchoscopic navigation did not improve the diagnosis rate of pulmonary peripheral lesions, but shortened the time to locate the lesions.
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Objective To explore the diagnostic value of radial ultrasound (EBUS) combined with virtual navigation (VBN) in peripheral pulmonary nodules.Methods Two hundreds and forty cases of peripheral pulmonary nodules(0.8 cm≤diameter≤3 cm) in the respiratory department of our hospital from July 2014 to July 2015 were included and according to the different guide de vices,which were divided into the radial ultrasound combined with virtual navigation group (EBUS+ VBN group),radial ultrasound group (EBUS group),virtual navigation group (VBN group) and control group.The diagnostic rates were compared among the four groups and among different sizes of lesion.The time of lesion location and operating time were also compared between the EBUS+ VBN group and EBUS group.Results Among 240 cases,the diagnostic rate in the EBUS+VBN group was highest(81.67％),and the diagnostic rates had statistically significant difference among the four groups (x2=19.344,P=0.00);the diagnostic rates of lesions less than 2 cm in diameter were lower than that of lesions ＞2 cm in the EBUS+-VBN group and EBUS group,but without statistically significant difference (x2 =2.04,3.40,P =0.15,0.07);the locating lesions time and operating times in the EBUS+ VBN group were shorter than those in the EBUS group,but the difference between them was not statistically significant (P=0.03,0.04).Conclusion EBUS combined with VBN could improve the diagnostic rate of peripheral pulmonary nodules and shorten the time of lesion location and operating time.
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Objective To explore the influence of polyamine-cholesterol cationic liposome (PCL)-mediated CpGODN aerosol on eosinophiles in the lung tissue of mouse asthma model. Methods Mouse asthma model was replicated by challenging with 1% ovalbumin aerosol. Mice were categonied into four groups, namely normal control, asthma control, CpGODN/PCL treatment group and CpGODN treatment group (6 each). The left lungs of mice were harvested, serially sectioned, hematoxylin and eosin (HE)-stained, and the infiltration of eosinophiles (EOS) was examined under microscope. Meanwhile, the bronchoalveolar lavage fluid (BALF) was collected for total and eosinophil cells count. Results An ovalbumin challenged mouse asthma model was successfully replicated. Pathological observation of the lung of asthma control showed increase in mucous secretion in alveolar space and peribronchial infiltration of large amount of inflammatory cells, primarily EOS and lymphocytes. The total cell number, EOS number and the ratio in BALF were significantly higher in asthma control group compared with that in both normal control group and CpGODN treatment group (P
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Objective To evaluate the transfection efficiency and cytotoxicity of polyamine cationic liposomes (PCL), and to screen for a cationic liposome with high potency and low toxicity. Methods Four kinds of PCLs, namely PCL-A, PCL-B, PCL-C and PCL-D, were prepared with polyamine cationic lipid TC-Chol or DC-Chol and the neutral phospholipid DOPE in different molar ratios (1∶1 or 3∶1). The morphology of the liposome was observed by transmission electron microscope. Plasmid PIRES2-EGFP, which contained an enhanced green fluorescencent-protein (EGFP) reporter gene, was transfected into Hela cells or Hep2 cells with PCL-A, PCL-B, PCL-C and PCL-D, respectively. Expression of EGFP was examined by fluorescence microscopy and the cytotoxicity of the transfected cells was determined with MTT assay. Results The shape of PCL-A, PCL-B, PCL-C and PCL-D was round or oval, most of the PCLs were about 50-200nm in diameter. The transfection efficiency of the four kinds of PCLs was about 19.8%-42.1%. Among them, PCL-C prepared with TC-Chol and DOPE at a molar ratio of 3∶1 showed a higher transfection efficiency (39.5%-42.1%) and lower cytotoxicity (cellular survival rate was 86.57%?3.23%-84.43%?4.18%). Conclusions The polyamine cationic liposome prepared with polyamine cationic lipids TC-Chol or DC-Chol and the neutral phospholipid DOPE at a certain molar ratio mediated the transfection of the plasmid PIRES2-EGFP into mammalian cells. PCL-C exhibited higher transfection efficiency and lower cytotoxicity, and it might be a promising agent for gene transfection and gene therapy.