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Objective:To investigate whether the hypervirulent Klebsiella pneumoniae (hvKP) induces liver abscess through activating NLRP3 inflammasome. Methods:K1-hvKP and K35-non-hvKP bacterial suspensions were intraperitoneally injected into C57BL/6 mice to establish the models of liver abscess. Human peripheral blood neutrophils were sorted by immunomagnetic beads with CD45 + and Gr-1 + , and the purity was detected by flow cytometry. The concentrations of capsular polysaccharide of K1-hvKP and K35-non-hvKP were detected by total carbohydrate assay kit. The expression of IL-18 and IL-33 by neutrophils at mRNA and protein levels was detected by real-time fluorescence quantitative PCR and ELISA, respectively. The activation of NLRP3 inflammasome in neutrophils was detected by Western blot. Neutrophil extracellular trap formation (NETosis) was observed under confocal laser scanning microscope. Results:The C57BL/6 mice with K1-hvKP infection had significantly serious liver abscess as compared with the K35-non-hvKP-infected mice. The purity of human neutrophils was more than 95%. The concentration of capsular polysaccharide in K1-hvKP was significantly higher than that in K35-non-hvKP. Compared with K35-non-hvKP, K1-hvKP significantly promoted the neutrophils to express IL-18 and IL-33 at both mRNA and protein levels, enhanced the activation of NLRP3 and induced NETosis.Conclusions:This study suggested that hvKP could promote NETosis by activating NLRP3 inflammasome to cause liver abscess.
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Objective:To investigate the role of down-regulating serine racemase (SRR) in alleviating the β-amyloid peptide (Aβ) induced neurotoxicity and synaptic damage and possible mechanism in PC12 cells.Methods:(1) PC12 cells cultured in vitro were divided into 0, 20, 40 and 80 μmol/L Aβ 25-35 treatment groups; they were treated with 0, 20, 40 and 80 μmol/L Aβ 25-35 for 24 h, respectively; cell counting kit (CCK)-8 was used to detect the survival rate of cells in each group, and Western blotting was used to detect the SRR protein expression. PC12 cells were treated with 40 μmol/L Aβ 25-35 for 0, 12, 24 and 48 h, respectively; cell survival and SRR protein expression were detected by CCK-8 and Western blotting, respectively. (2) PC12 cells were divided into control group, nonsense sequence group, SRR small interfering RNA (siRNA) group 1, SRR siRNA group 2, and SRR siRNA group 3; cells in the later three groups were transfected with SRR nonsense sequence or different SRR siRNA sequences, respectively; 48 h after that, Western blotting was used to detect the SRR protein expression of cells in each group, and SRR siRNA with best effect was selected for subsequent experiments. (3) PC12 cells were divided into control group, AD group, AD+nonsense sequence group, and AD+SRR siRNA group; cells in the latter two groups were transfected with nonsense sequence or SRR siRNA for 48 h, respectively; cells in the latter three groups were added 40 μmol/L Aβ 25-35, and cells in the control group were added same amount of solvent; 24 h after treatment, the SRR protein expression was detected by Western blotting, cell survival was detected by CCK-8, cell apoptosis was detected by Hoechst 33258 fluorescent staining, Caspase 3 activity was detected by enzyme linked immunosorbent assay, and the expressions of activated Caspase 3, N-methyl- D aspartate (NMDA) receptor-associated proteins and postsynaptic dense protein 95 (PSD95) were detected by Western blotting. Results:(1) The survival rate of cells in 0, 20, 40 and 80 μmol/L Aβ 25-35 treatment groups was successively decreased and the SRR protein expression was successively increased, with significant differences ( P<0.05); PC12 cells treated with 40 μmol/L Aβ 25-35 for 0, 12, 24 and 48 h had successively decreased survival rate and successively increased SRR protein expression, with significant differences ( P<0.05). (2) The SRR protein expressions in the SRR siRNA group 1, SRR siRNA group 2 and SRR siRNA3 group 3 were significantly decreased as compared with those in the control group and nonsense sequence group ( P<0.05), and the decrease in the SRR siRNA group 2 was the most obvious. (3) As compared with the control group, the cells in the AD group had significantly increased SRR protein expression and apoptosis rate, statistically decreased cell survival rate, significantly increased Caspase 3 activity and activated Caspase 3 protein expression, significantly increased protein expressions of NMDA receptor 2A (NMDAR2A) and NMDA receptor 2B(NMDAR2B), and statistically decreased PSD95 protein expression ( P<0.05); as compared with cells in the AD group, cells in the AD+SRR siRNA group had significantly decreased SRR protein expression and apoptosis rate, statistically increased cell survival rate, significantly decreased Caspase 3 activity and activated Caspase 3 protein expression, significantly decreased NMDAR2A protein expression, and statistically increased PSD95 protein expression ( P<0.05). Conclusion:Down-regulation of SRR expression can reduce the NMDAR2A protein expression, alleviate the over-activation of NMDA receptor, reduce the cell apoptosis, improve cell survival rate, protect nerve cells, increase PSD95 protein expression, and alleviate synaptic damage in PC12 cells.
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Objective To investigate the clinical and pathological features of ischemia renal injury.Methods Patients with biopsy-proven ischemia renal injury in the Department of Nephrology,Peking University First Hospital from 2010 to 2018 were retrospectively enrolled in the present study.The demographic information and laboratory data were collected.And the severity of pathologic changes including glomeruli,arteriole and tubulo-interstitial fibrosis (IFTA) were semiquantitatively scored.Arterioles with a ratio of inner/outer luminal diameter greater than 0.5 without hyalinosis were diagnosed as normal ones.The relationships between estimated glomerular filtration rate (eGFR),urine protein excretion and pathological changes were analyzed.Results A total of 52 patients were enrolled in the study,including 39 males (75.0%).The age of the patients was (45.0±12.7) years at biopsy.Among them,50 patients (96.2%) had a history of hypertension with a median duration of 66 (24,138) months.Forty-one patients (78.9%) were overweight or obese.The median urinary protein excretion was 0.75 (0.27,1.32) g/d with 3 cases over 3 g/d.Fifteen patients (28.8%)presented with microhematuria and twenty-seven patients (51.9%) with eGFR lower than 60 ml· min-1·(1.73 m2)-1,respectively.The ratio of arteriolar inner/outer luminal diameter was 0.43+0.05 and the percentage of normal arterioles was 29.0%+17.0%.There were 21 patients (40.4%) found with arteriole hyalinosis.The ratio of arteriolar inner/outer luminal diameter correlated with the percentage of glomerular lesions (rs=-0.312,P=0.024),the semiquantitative scale of IFTA (rs=-0.291,P=0.037) and eGFR (r=0.339,P=0.014),respectively.Hypertension duration and body mass index (BMI) showed a negative correlation with the ratio of arteriolar inner/outer luminal diameter (rs=-0.303,P=0.029 and rs=-0.274,P=0.050,respectively) and a positive correlation with serum complement 3 level (rs=0.358,P=0.020 and rs=0.432,P=0.004,respectively).Conclusions Renal ischemia injury may be found in young and middle-aged patients.The characteristic of clinical features is mild to moderate proteinuria accompanied with a certain degree of eGFR decline,while a small number of patients may have microhematuria and marked proteinuria.The ratio of arteriolar inner/outer luminal diameter has a negative correlation with the percentage of glomerular lesions and the semiquantitative scale of IFTA and a positive correlation with eGFR,respectively.Hypertension and obesity are closely related to vascular lesions.
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Objective@#To investigate the clinical and pathological features of ischemia renal injury.@*Methods@#Patients with biopsy-proven ischemia renal injury in the Department of Nephrology, Peking University First Hospital from 2010 to 2018 were retrospectively enrolled in the present study. The demographic information and laboratory data were collected. And the severity of pathologic changes including glomeruli, arteriole and tubulo-interstitial fibrosis (IFTA) were semi-quantitatively scored. Arterioles with a ratio of inner/outer luminal diameter greater than 0.5 without hyalinosis were diagnosed as normal ones. The relationships between estimated glomerular filtration rate (eGFR), urine protein excretion and pathological changes were analyzed.@*Results@#A total of 52 patients were enrolled in the study, including 39 males (75.0%). The age of the patients was (45.0±12.7) years at biopsy. Among them, 50 patients (96.2%) had a history of hypertension with a median duration of 66 (24, 138) months. Forty-one patients (78.9%) were overweight or obese. The median urinary protein excretion was 0.75 (0.27, 1.32) g/d with 3 cases over 3 g/d. Fifteen patients (28.8%) presented with microhematuria and twenty-seven patients (51.9%) with eGFR lower than 60 ml·min-1·(1.73 m2)-1, respectively. The ratio of arteriolar inner/outer luminal diameter was 0.43±0.05 and the percentage of normal arterioles was 29.0%±17.0%. There were 21 patients (40.4%) found with arteriole hyalinosis. The ratio of arteriolar inner/outer luminal diameter correlated with the percentage of glomerular lesions (rs=-0.312, P=0.024), the semiquantitative scale of IFTA (rs=-0.291, P=0.037) and eGFR (r=0.339, P=0.014), respectively. Hypertension duration and body mass index (BMI) showed a negative correlation with the ratio of arteriolar inner/outer luminal diameter (rs=-0.303, P=0.029 and rs=-0.274, P=0.050, respectively) and a positive correlation with serum complement 3 level (rs=0.358, P=0.020 and rs=0.432, P=0.004, respectively).@*Conclusions@#Renal ischemia injury may be found in young and middle-aged patients. The characteristic of clinical features is mild to moderate proteinuria accompanied with a certain degree of eGFR decline, while a small number of patients may have microhematuria and marked proteinuria. The ratio of arteriolar inner/outer luminal diameter has a negative correlation with the percentage of glomerular lesions and the semiquantitative scale of IFTA and a positive correlation with eGFR, respectively. Hypertension and obesity are closely related to vascular lesions.
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Objective To investigate the mechanism of linezolid resistance in methicillin-resistant Staphylococcus epidermidis ( MRSE) strains isolated in Hangzhou area. Methods Twenty-three linezolid-re-sistant Staphylococcus epidermidis ( LRSE) strains were isolated from the patients with septicaemia, urinary tract infection or infectious pleuritis in several hospitals in Hangzhou area during May, 2013 to April, 2015. The minimal inhibition concentrations ( MICs) of thirteen different antibiotics against the LRSE strains were detected by using E-test. Pulsed-field gel electrophoresis ( PFGE) and cluster analysis were performed for homology analysis. Correlations between linezolid resistance in the LRSE strains and G2576T mutation at theⅤ functional region of 23S rRNA gene or cfr gene were determined by PCR and sequencing analysis. Re-sults The MICs of linezolid to 15 LRSE strains were higher than 256 μg/ml while to the other 8 LRSE strains were 6 or 8 μg/ml. All of the 23 LRSE strains were resistant to both oxacillin and cefoxitin. Among the LRSE strains with high linezolid MICs (MIC>256 μg/ml), LRSE1 and LRSE2, LRSE3-LRSE6 and LRSE9-LRSE12 respectively belonged to the same clone line and came from the same hospital. All of the 23 LRSE strains carried the cfr gene. Moreover, the G2576T mutation at theⅤfunctional region of 23S rRNA gene was detected in the 15 LRSE strains with high linezolid MICs ( MIC>256 μg/ml ) , but not in the 8 strains with lower linezolid MICs ( MIC=6 or 8 μg/ml) . Conclusion There are significant differences in linezolid resistance among LRSE strains isolated in Hangzhou area. The LRSE strains are methicillin-resist-ant coagulase negative Staphylococcus ( MRCNS) strains and widely distributed. The linezolid resistance in LRSE strains is related to the G2576T mutation at theⅤfunctional region of 23S rRNA gene and cfr gene.
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<p><b>OBJECTIVE</b>To investigate the sensitivity and specificity of eosin-5'-maleimide (EMA)assay for the diagnosis of hereditary spherocytosis (HS), and to verify the stability of reagent and samples.</p><p><b>METHODS</b>EMA flow cytometry test, NaCl-osmotic fragility test and acidified glycerol lysis test were performed using peripheral blood samples from 80 patients with HS and 44 patients with other blood diseases, the sensitivity and specificity of the three methods were compared, and the feasibility of EMA binding test was estimated. The stability of EMA reagent and HS samples stored at different temperatures were tested.</p><p><b>RESULTS</b>Among the 124 tested samples, the sensitivity and specificity of EMA binding test was 0.925 and 0.954, that of NaCl-osmotic fragility test was 0.950 and 0.455, and that of acidified glycerol lysis test was 1.000 and 0.318, respectively. Although the sensitivity of NaCl-osmotic fragility test and acidified glycerol lysis test was a little higher than that of EMA binding test, the specificity of the former two methods was poor, they couldn't clearly distinguish whether spherocytosis is hereditary spherocytosis. The experiment results showed that EMA was sensitive to the temperature and should not be stored in a small aliquots at -80 ℃ over a period of 6 months. The stability of the HS sample was better, 6 days storage at 4 ℃ and 3 days storage at room temperature had no influence on the results.</p><p><b>CONCLUSION</b>EMA binding test by flow cytometry showed good sensitivity and specificity for HS diagnosis. EMA reagent should be stored at-80 ℃ and the HS samples should be tested within 6 days storage at 4 ℃ and 3 days at room temperature.</p>
الموضوعات
Humans , Ankyrins , Blood , Eosine Yellowish-(YS) , Flow Cytometry , Hematologic Tests , Sensitivity and Specificity , Spherocytosis, Hereditary , Blood , Diagnosisالملخص
<p><b>OBJECTIVE</b>To investigate the myeloperoxidase (cMPO) expression pattern by flow cytometry (FCM) in patients with acute myeloid leukemia (AML) and its role in classifying AML.</p><p><b>METHODS</b>Eight- color multiparametric FCM with CD45/SSC gating was used to determine the cMPO expression in 502 AML patients.</p><p><b>RESULTS</b>The positive rate of cMPO in all patients was 58.0%, in which the proportion of normal positivity, dim positivity and partial positivity was 21.5%, 34.1% and 2.4%, respectively. The remaining case (42.0%) were all negative. In AML with t (15;17)(q22;q12)/PMLRARα, the positive rate was the highest (100%) and the intensity was similar to that of the normal granular leukocytes, followed by AML with t (8;21(q22;q22/RUNX1-RUNX1T1, the positive rate was 91.4% and the intensity was mostly dim. AML with minimal differentiation and acute megakaryoblastic leukemia were all cMPO negative. The positive rates of cMPO in the remaining subtypes were between 22.7% and 76.2%.</p><p><b>CONCLUSION</b>The positive rate and intensity of cMPO were significantly different among different subtypes of AML.</p>
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Humans , Cell Differentiation , Flow Cytometry , Granulocytes , Leukemia, Myeloid, Acute , Classification , Genetics , Peroxidase , Geneticsالملخص
OBJECTIVE@#The purpose of this study is to analyze the development of musicality in children after cochlear implantation, and provide a clinical database for the evaluation of their musicality.@*METHOD@#Twenty-six children with cochlear implants (CI group) participated in this research. They received cochlear implants at the age of 11 to 68 months with a mean of 35.6 months. Seventy-six infants as a control group aged from 1 to 24 months with a mean of 6.1 months participated in this study, whose hearing were considered normal by passing the case history collection, high-risk registers for hearing loss and hearing screening using DPOAE. The music and young children with CIs: Musicality Rating Scale was used to evaluate their musicality. The evaluation was performed before cochlear implantation and 1, 3, 6, 9, 12, 24 months after cochlear implantation for children with cochlear implants. The evaluation was also performed at 1, 3, 6, 9, 12, 24 months for children with normal hearing.@*RESULT@#The mean scores of musicality showed significant improvements with time of CI use for CI group (P0.05). Significant difference was noted between the two groups at 24 months (P<0.05).@*CONCLUSION@#The musicality of children with cochlear implants improved significantly with time after cochlear implantation. The most rapid growth was found in the first year after cochlear implantation.
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Child, Preschool , Female , Humans , Infant , Male , Child Development , Cochlear Implantation , Music , Postoperative Periodالملخص
With the development of bio-technological drugs, drug immunogenicity evaluation has become key factor of clarifying safety and efficacy of these drugs. It has become the focus to establish a stable and reliable evaluation system. Due to the advantages such as continuous real-time monitoring, surface plasmon resonance (SPR) technology has been widely used in bio-technological drugs immunogenicity assessments. Our study applied this technology to detect anti-drug antibody (ADA) of a recombinant human anti-rabies monoclonal antibody NM57 in the sera of 48 volunteers admitted in phase I clinical trials. This method could satisfy the basic requirements of detection of ADA.
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OBJECTIVE@#Audiometric configuration distribution of hearing loss was analyzed for public policy efforts of hearing loss prevention and rehabilitation.@*METHOD@#According to the archived records of the Clinical Audiology Center of Beijing Tongren Hospital, Capital Medical University, audiometric configuration distribution was analyzed by different type,degree of hearing loss and age factor.@*RESULT@#1. Overall audiometric configuration distribution: the percentages of "sloping", "rising", "flat", "U-shaped", and other type of audiometric configuration distributions are 52.8%, 7.6%, 15.4%, 13.2% and 11.0%, respectively. No difference was found between male and female in the audiometric configuration distribution; 2. Audiometric configuration distribution by type of hearing loss: sloping hearing loss dominants sensorineural and mixed hearing loss; 3. Audiometric configuration distribution by degree of hearing loss: sloping hearing loss dominants mild, moderate and severe hearing loss, U-shaped hearing loss dominants profound hearing loss; 4. Audiometric configuration distribution by age: for age group 5 to 7, the highest prevalence of audiometric configuration is "sloping", followed by "flat", "U-shaped", other type and "rising"; with the age increasing, the percentage of sloping hearing loss increased dramatically.@*CONCLUSION@#Audiometric configuration distribution shows obvious characteristic of on the type, degree of hearing loss and age factor. "sloping" hearing loss dominates the audiometric configuration.
الموضوعات
Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Age Factors , Audiometry , Audiometry, Pure-Tone , Auditory Threshold , Hearing Lossالملخص
<p><b>BACKGROUND</b>Because invasion and metastasis of cancer threaten seriously human's life, it is of more important clinical significance to understand and evaluate the biologic behavior of can-(cer) correctly. Abnormal expression of E-cadherin and β-catenin plays important roles in invasion and metastasis of cancer. The objective of this study is to investigate their expression in non-small cell lung cancer (NSCLC) and to find out their correlation with histological type, cell differentiation, metastasis and prognosis of NSCLC.</p><p><b>METHODS</b>The expression of E-cadherin and β-catenin was detected in 129 NSCLC tissues by high sensitive S-P immunohistochemical method.</p><p><b>RESULTS</b>The abnormal expression rate of E-cadherin and β-catenin was 62.0% and 65.1% respectively. The abnormal expression rate of E-cadherin in squamous cell carcinoma was much higher than that in adenocarcinoma (P < 0.05). The abnormal expression rate of E-cadherin and β-catenin in poorly differentiated cells was significantly higher than that in well and moderately differentiated cells (P < 0.05, P < 0.05). Stage III/IV NSCLC showed markedly higher abnormal expression rate of E-cadherin and β-catenin than stage I/II NSCLC did (P < 0.01, P < 0.01). The abnormal expression rate of E-cadherin and β-catenin in patients with lymphatic metastasis was significantly higher than that in those without lymphatic metastasis (P < 0.05, P < 0.01). The mean survival duration and 5-year survival rate in patients with normal E-cadherin and β-catenin expression were remarkably higher than those in patients with abnormal expression (P < 0.05, P < 0.01).</p><p><b>CONCLUSIONS</b>The abnormal expression of E-cadherin and β-catenin is closely related to histological type, differentiation and metastasis in NSCLC. Detection of their expression might be helpful to predict prognosis of NSCLC.</p>