1.Protective effect and mechanism of ginsenoside Rg1 on H2O2induced hippocampal neurons aging due to down-regulate NOX2 mediated NLRP1 inflammasome activation in vitro
Tan-Zhen XU ; Ya-Li CHEN ; Xiao-Yan SHEN ; Ling-Ling SUN ; Bi-Qiong ZHANG ; Da-Ke HUANG ; Wei-Zu LI
Chinese Journal of Pharmacology and Toxicology 2018;32(4):321-321
OBJECTIVE To explore the protective effects and mechanisms of Ginsenoside Rg1 (Rg1) on H2O2-induced hippocampal neurons aging in vitro. METHODS The primary culture hippo-campal neurons(7 d)were randomly placed into six groups:normal control group,H2O2(200 μM)treat-ment group,and H2O2+Rg1(1,5 and 10μM)groups.The neurons were with Rg1(1,5 and 10 μmol·L-1) for 6h. H2O2(200 μmol·L-1) was added to the medium and incubate for 18 h. The Dihydroethidium (DHE) staining was performed for ROS production assessment. The LDH release and Hoechst 33258 were performed to examine the neuronal damage and apoptosis. The immunoblot was used to deter-mine the expression of β-Gal,NOX2,p22phox,p47phox,NLRP-1,ASC and Caspase-1 in hippocampal neurons.The ELISA was performed to detect the levels of IL-1β and IL-18 released in the supernatant in hippocampal neurons.RESULTS Rg1(5 and 10 μmol·L-1)significantly reduced the ROS production, attenuated H2O2-induced neuronal damage and apoptosis (P<0.05, P<0.01). The immunoblot results showed that Rg1(5 and 10 μmol·L-1)treatment significantly decreased the expression of β-Gal,NOX2, p22phox,p47phox,NLRP-1,ASC and Caspase-1 in hippocampal neurons(P<0.05,P<0.01).Additionally, Rg1(5 and 10 μmol·L-1)treatment significantly decreased IL-1β and IL-18 release in the supernatant. CONCLUSION The protective effect of Rg1 in H2O2-induced hippocampal neurons aging may be due to inhibit NOX2-NLRP1 activation.
2.Study on fluoroquinolone resistance and the relationship between resistance and mutations of gyrA and parC in Neisseria gonorrhoeae.
Jin-shui XU ; Bei WANG ; Chang-xian WANG ; Hai-jian GOU ; Zu-huang MI ; Jian-hua LING
Chinese Journal of Epidemiology 2006;27(8):702-704
OBJECTIVETo study the phenotypic and genotypic resistance to Fluoroquinolones in Neisseria gonorrhoeae (NG) isolated in Jiangsu province of China.
METHODSIn-vitro, susceptibility testing of ciprofloxacin and levofloxacin against ninety-five clinical isolates were determined by agar dilution method. Detection of mutation in the gyrA and parC genes was performed by polymerase chain reaction (PCR) assay and sequence analysis.
RESULTSThe clinical isolates demonstrated 100% resistance to ciprofloxacin. Based on gyrA and parC mutations, 18 types could be categorized among the 54 isolates. Based on the same gyrA mutations,isolates with high MIC appeared to have had more mutations in parC gene.
CONCLUSIONThe status of resistance to ciprofloxacin in NG was quite serious, and ciprofloxacin treatment for the treatment of NG infections in Jiangsu province should not be recommended. The results from this study suggested that mutations in the parC gene had contributed to the development of high Fluoroquinolone resistance in NG.
China ; DNA Gyrase ; genetics ; DNA Topoisomerase IV ; genetics ; Drug Resistance, Bacterial ; Fluoroquinolones ; pharmacology ; Genotype ; Gonorrhea ; drug therapy ; Humans ; Neisseria gonorrhoeae ; drug effects ; genetics ; isolation & purification ; Phenotype
3.Activated changes of platelet ultra microstructure and plasma granule membrane protein 140 in patients with non-small cell lung cancer.
Yi ZHUGE ; Jian-ying ZHOU ; Guang-die YANG ; De-ling ZU ; Xiao-liang XU ; Ming-qing TIAN ; Guo-hua LU
Chinese Medical Journal 2009;122(9):1026-1031
BACKGROUNDNon-small cell lung cancer (NSCLC) is the leading cause of cancer mortality worldwide. Platelet activation may play an important role in pathologic progress in lung cancer. In this study, we aimed to clarify the influence of activated platelets on lung cancer generation and growth, and the relationship among these functional and ultrastructural changes of platelets and the severity of pathogenetic condition in these patients with NSCLC.
METHODSOne hundred and thirty-six cases of patients with pathologically confirmed NSCLC were included in this study. Fifty-four healthy people were enrolled as controls. The change of ultra microstructure and activity of blood platelets were observed under the transmission and scanning electron microscope. Simultaneous determination of plasma granule membrane protein 140 (GMP-140) was made.
RESULTSTransmission electron microscopy showed remarkable changes of ultra microstructure of platelets in patients with NSCLC, including swelling, increase of a-granules, vesicles, and glycogenosome. Scanning electron microscopy showed many more surface processes and wrinkles on platelets in patients with NSCLC. The reference plasma levels of GMP-140 of healthy controls were (18.2 +/- 2.7) microg/L. The plasma levels of GMP-140 in patients with NSCLC were (47.8 +/- 12.3) microg/L, which were much higher than those of the controls. There was a medium positive correlation between plasma levels of GMP-140 and amount of a-granules (r = 0.514, P < 0.01) and a high positive correlation between plasma levels of GMP-140 and area of platelet (r = 0.84, P < 0.01) in patients with NSCLC. The Kaplan-Meier survival curve analysis showed significant shift to the left in patients with NSCLC whose a-granules per platelet were 19 or more compared to those 18 or less (Log rank statistic, chi(2) = 17.38, P < 0.01).
CONCLUSIONSThere are significant activated changes of ultra microstructure and increased activity of blood platelets in patients with NSCLC. These activated platelets may play an important role in the generation and growth of lung cancer. These changes can be used as a diagnostic index of severity, progression, and prognosis of NSCLC.
Adult ; Aged ; Blood Platelets ; ultrastructure ; Carcinoma, Non-Small-Cell Lung ; blood ; drug therapy ; mortality ; ultrastructure ; Female ; Humans ; Male ; Microscopy, Electron, Transmission ; Middle Aged ; P-Selectin ; blood ; Survival Analysis
4.Study on reliability and validity of the Tinnitus Evaluation Questionnaire.
Peng LIU ; Gui-li XU ; Ming LI ; Zu-wei CAO ; En-qin GUO ; Ling-ling AN ; Nai-ping WANG ; Jian-ning ZHANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2012;47(9):716-719
OBJECTIVETo investigate the value of the Tinnitus Evaluation Questionnaire (TEQ) in clinical application.
METHODSCronbach's α coefficient was used to examine the reliability of the TEQ internal consistency. Examined the re-measured reliability by the correlation coefficient by two doctors' 1 - 3 hours interval questionnaires' scores. And inspected criteria validity according to the correlation coefficient of the TEQ and Tinnitus Handicap Inventory (THI).
RESULTSIn the 202 tinnitus patients, the TEQ Cronbach's α coefficient was 0.76 and re-measured reliability was 0.938. The THI correlation coefficient was 0.769. Among which, 99 patients feel tinnitus alleviated obviously after the treatment, the TEQ scores were significantly lower than that before the treatment (t = 21.42, P < 0.001).
CONCLUSIONSThe TEQ reflects the severity of tinnitus completely, and has preferable reliability and validity. The characteristics are concise, practical and exact. It is worthy of clinical application.
Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Reproducibility of Results ; Surveys and Questionnaires ; Tinnitus ; diagnosis ; Young Adult
5.Midterm outcomes of percutaneous transluminal septal myocardial ablation in patients with hypertrophic obstructive cardiomyopathy refractory to medication.
Shao-liang CHEN ; Fei YE ; Zu-ling XU ; Song LIN ; Bao-xiang DUAN ; Zhen-ling DAI ; Shou-jie SHAN ; Jun-jie ZHANG
Chinese Medical Journal 2006;119(13):1121-1124
Adult
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Aged
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Bundle-Branch Block
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etiology
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Cardiomyopathy, Hypertrophic
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surgery
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Catheter Ablation
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adverse effects
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methods
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Female
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Heart Septum
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surgery
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Humans
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Male
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Middle Aged
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Postoperative Complications
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etiology
6.Value of protein array in the diagnosis of Helicobactor pylori infection in children.
Xiao-Li SHU ; Hai-Ying CHENG ; Ling-Rong YANG ; Xu-Ping ZHANG ; Yan-Yi ZHANG ; Mi-Zu JIANG
Chinese Journal of Contemporary Pediatrics 2011;13(9):728-731
OBJECTIVETo study the value of multiple Helicobacter pylori (H.pylori) antibody detection by protein array in the diagnosis of H.pylori infection in children.
METHODSBiopsy specimens obtained by gastroscopy from 120 children with digestive system symptoms were detected by rapid urease test (RUT) and modified Giemsa staining. Positivity in both RUT and Giemsa staining was the "gold criterion" of H.pylori infection. Serum samples of these patients were obtained and the antibodies against cytotoxin associated gene A protein (CagA), vacuolating toxin A (VacA), urease, heat shock protein 60 (Hsp60) and RdxA (nitroreductase) were detected by protein array technique.
RESULTSH.pylori infection was identified according to the "gold criterion" in 60 children. Compared with the "gold criterion", the goodness of fit and the coefficient of contingency in the diagnosis of H.pylori infection of the following four groups antibody detection were all statistically significant (P<0.001): anti-Ure antibody alone, anti-Ure antibody combined with anti-CagA antibody, anti-Ure antibody combined with anti-VacA antibody and anti-Ure antibody combined with anti-CagA and anti-VacA antibody. The sensitivity, specificity and accuracy of the detection of anti-Ure antibody combined with anti-CagA antibody for the diagnosis of H.pylori infection were 81.7%, 91.7% and 86.7%, respectively. The antibody detection showed a high positive predictive value (90.7%) and a high negative predictive value (83.3%).
CONCLUSIONSThe antibody detection by protein array, especially the detection of anti-Ure antibody combined with anti-CagA antibody, is valuable in the diagnosis of H.pylori infection.
Adolescent ; Antibodies, Bacterial ; blood ; Child ; Child, Preschool ; Female ; Helicobacter Infections ; diagnosis ; Helicobacter pylori ; immunology ; Humans ; Male ; Protein Array Analysis ; methods ; Sensitivity and Specificity
7.Histopathological changes in livers of patients with chronic severe hepatitis B.
Li-jie ZHANG ; Tai-ling WANG ; Xu-hua LIU ; Ke-jia ZU ; Xin MENG ; Yue WANG ; Zhong-ping DUAN
Chinese Journal of Hepatology 2007;15(5):323-325
OBJECTIVETo investigate the histopathological features in livers of chronic severe hepatitis B (CSHB) patients.
METHODSHistology of 42 livers was studied. HE, Masson, Sweet and D-PAS staining and cytokeratin 7, CD68 and proliferating cell nuclear antigen immuno-histochemical staining were used in the study.
RESULTSIn CSHB, the livers showed massive or submassive necrosis in a background of other histological changes of chronic hepatitis B. The characteristic pictures of these livers were necrosis of all the hepatocytes in some nodules, while in other nodules there were only patchy necroses of the parenchyma. In some other nodules the necrotic hepatocytes were all removed and only the scaffolding stroma remained. Meanwhile, regeneration of hepatocytes and bile ductules were also seen.
CONCLUSIONSThe liver histopathological changes in CSHB are identical, but not of the same degree as those of acute severe and subacute severe hepatitis B. In making differential diagnoses for liver aspiration biopsies of these patients, this fact should be kept in mind.
Adult ; Aged ; Female ; Hepatitis B, Chronic ; diagnosis ; pathology ; Humans ; Liver ; pathology ; Male ; Middle Aged ; Staining and Labeling ; Young Adult
8.Expression of human phosphodiesterase 3A gene using baculovirus expression system in insect cell.
Yong-Min YAN ; Wen-Rong XU ; Hai-Bo ZHU ; Ling ZHANG ; Ren-Ren FEI ; Xiao-Liang WANG ; Hui QIAN ; Shun-Zu HAO ; Wen-Bing WANG
Acta Pharmaceutica Sinica 2005;40(9):810-813
AIMTo investigate the expression of recombinant human phosphodiesterase 3A (HPDE3A) using baculovirus expression system in Tn cell line.
METHODSThe HPDE3A cDNA was recombined with baculovirus, and then the recombinant was transfected into Tn cell line. The expression of HPDE3A in Tn cell line was detected and identified by the RT-PCR, SDS-PAGE and Western blotting.
RESULTSThe recombinant HPDE3A protein was stably expressed in Tn cell line and detected by the distinct morphological changes of Tn cell, RT-PCR, SDS-PAGE and Western blotting using polyclonal antibody. The M(w) of the recombinant protein was about 120 kD.
CONCLUSIONRecombinant HPDE3A can be expressed in Tn cell line using the baculovirus expression system, and thus provided the basic material for studying its bioactivity and application in screening for HPDE3A inhibitor.
3',5'-Cyclic-AMP Phosphodiesterases ; genetics ; metabolism ; Animals ; Baculoviridae ; genetics ; Cell Line ; Cyclic Nucleotide Phosphodiesterases, Type 3 ; Electrophoresis, Polyacrylamide Gel ; Moths ; cytology ; enzymology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Transfection
9.Vacuum-assisted biopsy and wire localization for the diagnosis of non-palpable breast lesions.
Kun-Lun SU ; Hai-Bin XU ; Zu-Jian HU ; Jun-Ling HE ; Ou-Ou YANG ; Wang-Hua HU
Chinese Journal of Oncology 2010;32(6):472-475
OBJECTIVETo compare the effectiveness and accuracy of the use of vacuum-assisted biopsy (VAB) versus wire localization (WL) in the diagnosis of non-palpable breast lesions (NPBL).
METHODSNinety-seven consecutive women with NPBL were randomized into VAB group and WL group. All specimens were identified by mammography. The patients were requested to score the cosmetic appearance of their breast after operation, and a numerical rating scale was used to measure pain on the first postoperative day. Underestimation rates for atypical ductal hyperplasia (ADH) and ductal carcinoma in situ (DCIS) were recorded if open surgical biopsy revealed DCIS and invasive cancer, respectively. Clear margins were also recorded in the two groups.
RESULTSVAB and WL located all the NPBL successfully. In the VAB group, the specimen volume was smaller than that of the WL group (2.3 cm(3) vs. 18.4 cm(3), P = 0.03). Underestimation rates of ADH and DCIS in the VAB group were 16.7% and 11.1%, respectively. The diagnostic accordance rate of VAB was 97.9%, the false negative rate was 2.1%, and there was no false positive case. The means of the numerical rating pain scale were different in both groups (1.7 for VAB vs. 2.5 for WL, P = 0.02). When cosmetic results were taken into account, 40 VAB patients had excellent outcomes and 8 good outcomes, compared with 25 excellent and 24 good for the WL group. There were better cosmetic outcomes with the VAB procedure (P < 0.05).
CONCLUSIONVAB is highly reliable and may avoid diagnostic open surgery in the majority of patients with benign lesions. However, because of the underestimation of histologic diagnosis and tumor margin involvement, VAB can not be used to completely substitute wire localization.
Adult ; Biopsy, Needle ; instrumentation ; methods ; Breast ; pathology ; Breast Neoplasms ; diagnosis ; pathology ; Carcinoma in Situ ; diagnosis ; pathology ; Carcinoma, Ductal, Breast ; diagnosis ; pathology ; Diagnostic Errors ; Female ; Fibroadenoma ; diagnosis ; pathology ; Humans ; Hyperplasia ; Middle Aged ; Precancerous Conditions ; diagnosis ; pathology ; Stereotaxic Techniques ; instrumentation ; Vacuum
10.Application of nested PCR and sequencing technique to detect point mutations of the 23S rRNA gene of Mycoplasma pneumoniae.
De-li XIN ; Zu-huang MI ; Xu HAN ; Ling QIN ; Jing LI ; Xi-jie LIU ; Shao-jie MA ; An-cun HOU ; Gui LI
Chinese Journal of Pediatrics 2008;46(7):522-525
OBJECTIVETo establish a quick method to detect drug resistance of Mycoplasma pneumoniae (MP) and study the condition of drug resistance in MP infection.
METHODSMP 23S rRNA target gene in throat swab specimens from 200 patients with suspected MP infection was detected by using nested PCR and DNA sequencing. The result of 23S rRNA gene detection was confirmed by MP isolation and drug susceptibility test in vitro for reliability.
RESULTSOf the 200 clinical specimens, 64 were proved to be positive for MP through MP-IgM antibody, MP specific 16S rRNA nested PCR and MP isolation . The 23S rRNA gene was amplified and the gene sequence was compared with MP reference strain in Genbank, 26 were identical to the reference strain, 38 had a point mutation in 23S rRNA. Among them, 35 had A to G mutation at position 2063, 1 had A to C mutation at position 2063 and 2 had A to G mutation at position 2064, the percentage of drug resistance was 59.4%. The sensitivity of the gene detection method was 10(2) ccu/ml and it was confirmed to be reliable by MP isolation and drug susceptibility test.
CONCLUSIONSThe gene detection method could detect MP drug resistant gene directly from clinical specimen, which has the advantages of high specificity, high sensitivity and quickness. It is of great significance for diagnosis of MP infection because MP isolation is difficult and time-consuming.
Adolescent ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Genes, rRNA ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Mycoplasma pneumoniae ; genetics ; isolation & purification ; Point Mutation ; Polymerase Chain Reaction ; RNA, Bacterial ; genetics ; RNA, Ribosomal, 23S ; genetics