1.Establishment of mouse septic peritonitis model with low pathogenic bacteria
Tao LUO ; Wenxiang HUANG ; Lixian WU
Journal of Third Military Medical University 1984;0(01):-
0.05). Conclusion The application of SRFE can decrease MLD effectively when form mice septic peritonitis models, especially for those bacteria with low pathogenicity.
2.Prevalence of virulence genes in Streptococcus pneumoniae strains isolated from clinical patients
Yishan DONG ; Wenxiang HUANG ; Tao LUO ; Cheng ZHANG ; Lixian WU
Chinese Journal of Microbiology and Immunology 2009;29(2):177-180
Objective To investigate the prevalence of virulence genes(ply, pspA, nanA, lytA, psaA) among Streptococcus pneumoniae recently isolated from clinical patients. Methods The 133 strains were isolated from patients in three teaching hospitals in Chongqing from 2006 to 2008. Polymerase chain reaction was used to screen for virulence genes (ply, pspA, nanA, lytA, psaA). Results The positive rate of lytA, psaA, ply, nanA and pspA in 133 clinical isolates were 94.7%, 85.0%, 82.7%, 84. 2% and 60.2%, respectively. The positive rates of the lytA, psaA, ply, nanA and pspA genes in 87 common serotypes isolates was 100%, 87.4%, 86.2%, 89.7%, 67.8%, respectively. Conclusion The total positive rates of five virulence genes in the 133 clinical strains were high. The positive rates of five genes in the com-mon serotypes isolates were higher than those in the no-common serotypes. These genes are important virulence genes of Streptococcus pneumoniae. They could be candidates for protein vaccine of Streptococcus pneumoniae.
3.The therapeutic effects of survivin antisense nucleic acid combined with paclitaxel on subcutaneous xenograft mouse model of Balb/c
Lixian WU ; Lisen HUANG ; Ruijia CHEN ; Jue TIAN ; Fang KE
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(4):467-471
Objective To explore the therapeutic effects of combined application of survivin antisense nucleic acid and taxol in subcutaneous xenograft mouse model of Balb/c and to preliminarily investigate the mechanism of the anticancer effects.Methods The model of subcutaneous tumor was established by hypodermic injection of C26 cells into Bal b/c mice.The mice were then randomly divided into five groups through the internal tumor injection:the blank group (C),lipo2000 group (L),paclitaxel group (T),survivin antisense nucleic acid group (A),and survivin antisense nucleic acid combined with paclitaxel group (A+T).We observed tumor growth,determined cell apoptosis by TUNEL method,and detected the expression of survivin by Western blot.Results ① All treatment groups had T/C<60%,which was significantly different from that of group L (P <0.05);the intervention was proved effective in vivo .The tumor inhibition rate of mice tumor weight showed that there were significantly curative effects in groups T,A and A+ T compared with that in group C (P < 0.05 ).The antitumor activity of paclitaxel (tumor inhibition rate of 21.82%±0.84%)could be improved by more than 59% through combination therapy (tumor inhibition rate of 54.1 6% ± 0.32%)concerning inhibition of tumor weight growth.② TUNEL method detected apoptotic cells:The tumor cells hardly had apoptosis in the blank group while T group and A group had a certain number of apoptotic cells.The experiment results suggested that PTX could promote tumor cell apoptosis,and that not only A+T strengthened the effect in killing tumor cells,but also the synergy of both could influence tumor resistance and ultimately make the effect in promoting tumor cell apoptosis conspicuous.③ The expression of survivin protein:The results showed that the expression of survivin protein in group A + T was obviously decreased without the expression of β-actin affected;it did not change significantly in group C compared with group L.The ratio of the A-value in groups T,A and A+T was 0.895 ±0.01 1,0.704 ±0.121 and 0.345 ± 0.01 9,respectively.Analysis of variance t-test showed that the expression level in group A+T obviously differed from that in groups C,L,A and T (P <0.05).Conclusion The combined therapy of survivin antisense nucleic acid and taxol can promote tumor cell apoptosis by downregulating the expression of survivin protein,reduce the body’s resistance to drugs and create synergetic effects.
4.XN4 inhibits proliferation of AML cells by inducing oxidative DNA damage
Lixian WU ; Lisen HUANG ; Xianling CHEN ; Fang KE ; Ming ZHENG ; Jianhua XU
Chinese Pharmacological Bulletin 2014;(4):478-483
Aim To investigate the cytotoxicity of XN4 against AML cells, and the underlying mechanisms by which XN4 might induce DNA damage and apoptotic cell death through reactive oxygen species ( ROS ) . Methods The proliferation inhibition ratio of AML cells was measured by MTT. The level of extracellular ROS, DNA damage, cell cycle process and apoptosis were tested by flow cytometry ( FCM ) . Western blot was applied to test the expression of proteins. Results XN4 significantly inhibited the proliferation of HL-60 and KG1α with IC50 ( 2. 79 ± 0. 15 ) μmol · L-1 and (2. 76 ± 0. 20) μmol·L-1 respectively. XN4 signifi-cantly increased the generation of intracellular ROS, followed by inducing DNA damage and activating the ATM-γ-H2AX signaling, which led to increases of cells in the S phases of the cell cycle. Subsequently, XN4 induced apoptotic cell death through activation of caspase-3 and Parp. Moreover, the above effects were all reversed by the ROS scavenger N-acetylcysteine ( NAC ) . Conclusion XN4-induced DNA damage and cell apoptosis in AML cells are mediated via ROS generation.
5.Genotypic and phenotypic analysis of SLC26A4 gene in deaf patients of Chifeng area by whole gene sequencing strategy
Yongyi YUAN ; Pu DAI ; Deliang HUANG ; Xiuhui ZHU ; Qingwen ZHU ; Dongyang KANG ; Lixian LIU ; Guochun TENG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(05):-
OBJECTIVE To investigate the genetic causes of deaf patients in a special educational school of Chifeng city, Inner Mongolia by SLC26A4 whole gene sequencing. This study focused on analyzing mutations of coding sequence of SLC26A4 gene and their relevant phenotype. METHODS DNA were extracted from peripheral blood of 134 deaf patients of Chifeng special educational school and 100 normal hearing controls in Northern China. SLC26A4 gene mutation was analyzed by direct sequencing for its 20 coding exons. All individuals found with SLC26A4 mutation were given temporal bone CT scan, and those with confirmed enlarged vestibular aqueduct and/or other malformation of inner ear were then given further ultrasound scan of thyroid and thyroid hormone assays. RESULTS The sequencing results revealed 32 cases carried SLC26A4 mutation. Twenty-nine cases underwent temporal bone CT scan. Twentycases were confirmed to have malformation of inner ear by CT scan (eighteen were EVA, one was EVA and other inner ear malformation and one was Mondini Syndrome). The shape and function of thyroid were confirmed to be normal by ultrasound scan of thyroid and thyroid hormone assays in nineteen of these 20 patients except one who had cystoid change in the right side of thyroid. Twelve types of novel variants of SLC26A4 gene were found. CONCLUSION Byscreening SLC26A4 gene coupled with temporal bone CT scan ,we could determine genetic cause related to this gene up to 14.93 % of deaf patients in special educational school of Chifeng city. SLC26A4 is another common gene besides GJB2 that cause deafness in this area. The discovery of novel variants of SLC26A4 gene makes the mutational and polymorphic spectrum more plentiful in Chinese population.
6.Study on Immunological Components and Their Contents in Subcutaneous Exudate Induced by Cutaneous Scraping Method
Zhicong WU ; Shiya LIU ; Lianghui LI ; Jiejing LAI ; Lixian HUANG ; Hua CHEN ; Zaoyuan KUANG ; Biaoyan DU
Journal of Guangzhou University of Traditional Chinese Medicine 2017;34(2):209-212
Objective To investigate the changes of the immunological factors in subcutaneous exudate and blood components of the rats receiving cutaneous scraping method,and to compare the changes of skin histopathological features before and after cutaneous scraping under microscope.Methods SD rats were randomly divided into two groups,cutaneous-scraping group and non-cutaneous-scraping group.And then each group was divided into three subgroups.The observation indexes included the levels of interleukin (IL)-1β,IL-6 and interferon gamma (IFN-γ) in the blood and the skin,routine blood examination,and skin histopathological features.Results In cutaneous-scraping group,the number of white blood cells in the blood and the levels of IL-1 β and IFN-γ in skin tissues were increased (P < 0.05),the hemolysis rate was increased (P < 0.05).However,the levels of IL-1β,IL-6 and IFN-γin the blood showed no obvious changes.Under the microscope,severe skin edema,vascular congestion and dilatation,and infiltration of inflammatory cells were found in the skin after cutaneous scraping.Conclusion The cutaneous scraping method can activate the immune response rapidly,and the immunological components of the subcutaneous exudate after cutaneous scraping are helpful to the disease treatment.
7.Study on pathogenicity of putative virulence gene of Enterococcus faecium.
Lixian WU ; Wenxiang HUANG ; Guofu WANG ; Xiaoping SUN
Journal of Biomedical Engineering 2009;26(3):601-605
The presence of hyl gene in 364 PFGE clones of Enterococcus faecium was detected by colony hybridization under conditions of high stringency. The isogenic hyl-deficient mutant (* hyl) was constructed with suicide pTX4577 and screened by allelic replacement. Moreover, an in vitro study was made on the effect of hyl gene detection on the growth ability of hylgene detection on the mutant, and an in vivo study was made on the decrease of virulence in the mouse peritonitis model. The results showed, in the clinical isolates, the positive percentage of hyl gene was 32.8%, which was significantly higher than that (5.3%) in the non-clinical isolates. The * hyl was selected by kanamycin and identified by PCR, pulsed field gel electrophoresis (PFGE) and Southern blot. The experimental evidence indicated that the growth ability of * hyl was remarkably reduced in comparison with that of the wild-type strain. The percentage survival of mice in TX2466 groups was 0, while that of * hyl groups was 50% at the same inoculum in mouse peritonitis. The differences were significant. These data suggest that hyl gene in specific E. faecium strains may be enriched in determinants that make them more likely to cause clinical infections. Being important in the pathogenesis, hyl gene is probably a major virulence factor of Enterococcus faecium.
Animals
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Enterococcus faecium
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genetics
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pathogenicity
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Genes, Bacterial
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Gram-Positive Bacterial Infections
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microbiology
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Hyaluronoglucosaminidase
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genetics
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Male
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Mice
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Mice, Inbred ICR
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Mutation
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Peritonitis
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microbiology
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Virulence
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Virulence Factors
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genetics
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metabolism
8.A clinical studv of 49 cases of invasive pulmonary aspergillosis
Huayin LI ; Liping ZHU ; Huiping LI ; Yi HUANG ; Xin ZHOU ; Xiajun RONG ; Huili ZHU ; Lixian HE ; Weiwu DENG ; Xinhua WENG
Chinese Journal of Internal Medicine 2008;47(12):1017-1021
Objective studyring the proven and probable invasive pulmonary aspergillosis(IPA) eases of some hospitals in Shanghai to provide evidence fur the improvement of IPA clinical diagnosis and therapy.Methotis Fortv-nine IPA cases were retrospectively analyzed for demography data,host tactors,underlying conditions.chest CT,microorganism and histopathology examination,as well as therapy and clinical outcome.ResultsOf 49 subjects including 19(38.8%)proven and 30(61.2%)probable IPA,3 pailents(6.1%)had no host factors,25 patients(51.0%)had IPA associated host factors and underlying conditions.while 21 patients(42.9%)had uncertained fundamental diseases.Chest CT evaluation demonstrated that radiological lesions include nodules in 29 patients,patching in 15,mass in 12,consolidation in 10.cavitation in 34,Halo sign in 19,air bronchogram in 18,crescentic sign in 6,bilateral in 33 and multifocal lesions in 38.The yielding rate of fungus culture in sputum was 26.5%(13/49),and in bronchoalveolar lavage fluid was 66.7%(10/15).Eleven of thirty-six patients(30.6%)had positive results of serum galactomannan antigen tests.Nineteen of twenty-one patients(90.5%)were proven as IPA by lung histologic examinations.Aspergillus fumigatus was the most common pathogen 81.0%(17/21).The responding rate to initial anti-fugus therapy wag 50%(21/42).Conclusion Our study suggests that in IPA patients,bilateral,muhifocal and nodular lesion could be the most common radiological characteristic,while Halo and crescentic sign occar occasionally.Invasive technologies are more valuable to IPA diagnosis.
9.Multi-center clinical studies on pazufloxacin sodium chloride injection in treatment for acute bacterial infections
Jing YANG ; Gengzhi GE ; Tian XIA ; Baoyuan CHEN ; Xiaowen HAN ; Bing LIU ; Lixian HE ; Xiaoqiang DING ; Zhongyi HUANG ; Zhongmin QIU ; Jun MA ; Ce SHEN ; Niansong WANG
Chinese Journal of General Practitioners 2008;7(1):19-21
Objective To evaluate efficacy and safety of locally-produced pazufloxacin mesilate sodium chloride injection in the treatment of bacterial infections of respiratory and urinary tract.Methods A multi-center double-blind randomized controlled clinical trial was carried out to evaluate efficacy and safety of pazufloxacin mesilate sodium chloride in treatment for acute bacterial infection, as compared to those of levofloxacin hydrochloride and glucose injection as control treatment.A total of 244 patients with acute bacterial infection of respiratory and urinary tract were enrolled in the studies.120 in trial group and 120 in control group, with four withdrawals.Pazufloxacin mesilate and levofloxacin were administered intravenously by drip at a dose of 300 mg and 200 mg, every 12 hours for 7 to 14 days for trial and control groups, respectively.Resuits Overall efficacy of pazufloxacin mesilate was 77.0 percent and 93.5 percent in treatment for acute bacterial infections of respiratory and urinary tract.respectively, and that of levofloxacin was 80.6 percent and 89.6 percent, respectively.Overall bacterial clearance rate WaS 91.5 percent for pazufloxacin mesilate, 89.6 percent for respiratory tract infection and 94.1 percent for urinary tract infection, respectively.and 93.4 percent for levofloxacin, 97.3 percent for respiratory tract infection and 89.7 percent for urinary tract infection, respectively.No significant difference in adverse drug reactions between the two groups(P>0.05)was found, with 4.88 percent and 7.44 percent for trial and control groups, respectively.Conclusions Pazufloxacin mesilate sodium chloride injection produced locally is a safe and effective antibiotic in treatment for acute infections of respiratory and urinary tract.
10.Effect of hypotension factor on endotracheal tube cuff-induced damage to tracheal mucous mem-brane of rabbits
Wenhao WU ; Shuxian MA ; Lixian WANG ; Jiangli WU ; Yanjie YANG ; Zhangnan SUN ; Lining HUANG
Chinese Journal of Anesthesiology 2017;37(11):1402-1404
Objective To evaluate the effect of hypotension factor on endotracheal tube cuff-in-duced damage to tracheal mucous membrane of rabbits. Methods Eighty healthy rabbits of both sexes, aged 3.0-3.5 months, weighing 2.5-3.5 kg, were divided into 16 groups(n=5 each)according to the cuff pressure and mean arterial pressure(MAP): different cuff pressures when MAP did not decrease groups(C1M1group, C2M1group, C3M1group, C4M1group), different cuff pressures when MAP de-creased by 20% of the baseline value groups(C1M2group, C2M2group, C3M2group, C4M2group), dif-ferent cuff pressures when MAP decreased by 30% of the baseline value groups(C1M3group, C2M3group, C3M3group, C4M3group), and different cuff pressures when MAP decreased by 40% of the baseline value groups(C1M4group, C2M4group, C3M4group, C4M4group). Different cuff pressures were 0, 10, 20 and 30 cmH2O.At 2 h of tracheal intubation, the tracheas in the cuff-compressed area were harvested and sliced for examination of the pathological changes of tracheal mucous membrane after haematoxylin and eosin staining(with a light microscope), and the damage to tracheal mucous membrane was scored. Results When at the same low pressure(MAP decreased by 20%, 30% and 40% of the baseline value), the score of damage to tracheal mucous membrane increased with the increasing cuff pressures(P<0.05). When at the same cuff pressure(10, 20 and 30 cmH2O), the score of damage to tracheal mucous membrane in-creased with the increasing MAP(P<0.05). There was interaction between cuff pressure and MAP, F=2.034(P<0.05). Conclusion There is interaction between the effects of hypotension factor and endotra-cheal tube cuff factor on damage to tracheal mucous membrane; hypotension factor can aggravate endotra-cheal tube cuff-induced damage to tracheal mucous membrane of rabbits.