1.Detection of integron structure and sequence of drug resistance gene cassette in clinical multidrug resistant Shigella dysenteriae strains
He YAN ; Minhua ZONG ; Lei SHI ; Yamasaki SHINJI
Chinese Journal of Infectious Diseases 2010;28(5):273-277
Objective Clinical strains of Shigella dysenteriae isolated from eastern parts of India in 1988, 1995 and 2002 were examined for the presence of class 1, 2 and 3 integrons which closely related with drug resistance and the distribution of resistance gene cassette in order to clarify the influence of integron system on drug resistance of Shigella dysenteriae. Methods Susceptibility to antimicrobial agents was tested by the disk agar diffusion method. Class 1, 2 and 3 integron genes (intI) were screened by polymerase chain reaction (PCR) in 16 clinical strains with drug resistance.The variable regions of gene cassette of positive strains were sequenced. Results All 16 isolates were resistant to at least 4 agents including β-lactams, aminoglycosides, tetracyclines, sulfonamides,chloramphenicols and quinolones. Class 1 integron gene was detected in 13 strains and all isolates carried class 2 integron which indicated that strains with two integron structures were detected simultaneously and class 3 integron was not detected. Class 1 integron inserted gene cassette was mainly blaara30 -aadA 1 family, conferring resistance to β-lactamase, spectinomycin and streptomycins isolates carried class 2 integron were mainly dfrAl-satl genes cassettes conferring resistance to methoxybenzyl aminopyrimidine and streptothricin, while dfrA\-sat\-aadA\ genes were present only in 4 isolates. Conclusions These data indicate that class 2 integrons are widespread in Shigella dysenteriae strains, and closely associated with multidrug resistance of Shigella.
2.Embryo-fetus development toxicity of a novel PPAR-δ agonist in rat.
Hua-Yun GONG ; Yong ZHU ; Zong-He LI ; Xiao-Yan FAN ; Rong FAN ; Fang-Tong WANG
Acta Pharmaceutica Sinica 2014;49(11):1536-1542
The study aims to investigate the embryo-fetus development toxicity of the novel PPAR-δ agonist HS060098 on SD rats. The pregnant rats that were randomly divided into the solvent control group (1% hydroxypropyl methyl cellulose water solution) and HS060098 suspension groups (10, 30 and 100 mg x kg(-1) xd(-1)) were orally administered with HS060098 suspension or vehicle during the gestation of 6 -15 days (GD6-15). At termination (GD20), female rats were sacrificed. The pregnant females were evaluated by corpora lutea count, implantation sites, existence and death of embryos. Fetal sex, weight, externals, variations and malformations of viscus and skeleton were observed. The results show that there were no significant abnormality in maternal general conditions and fetal appearance as well as viscera, but in the 100 mg x kg(-1) x d(-1) group, the maternal weight gain decreased greatly (P < 0.01) and the skeletal ossification delayed remarkably (P < 0.01); in the 30 mg x kg(-1) xd(-1) group, the fatal and litter number of incompletely ossified sternebrae II was higher than those of the control group (P < 0.05); the skeletal malformations occurred in all dose groups, which indicate that the novel PPAR-δ agonist HS060098 had maternal toxicity and adversely effected fetal skeletal development under the experimental conditions.
Animals
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Bone and Bones
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drug effects
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Embryonic Development
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drug effects
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Female
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Fetal Weight
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PPAR delta
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agonists
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Pregnancy
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Rats
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Toxicity Tests
3.The effect of Polygonatum Sibiricum Polysaccharide on the expression of OPG and RANKL in the rat model of ovariectomy-induced osteoporosis
Fangna YAN ; Gaofeng ZENG ; Shaohui ZONG ; Xiaoming PENG ; Pingping WU ; Lei ZHANG ; Jichen HE ; Chengming WEI ; Xiongzhi SHI
The Journal of Practical Medicine 2017;33(8):1243-1246
Objective To investigate PSP on bone microstructures,Ca,P,OPG and RANKL of osteoporotic rat model.Methods Thirty female rats randomly divided into five groups:Sham,OVX,H-,M-,L-PSP.Sham and OVX were irrigated stomachsaline;PSP solution was gavaged to other groups.After 8-week,bone microstructures of tibial metaphyseal,Ca,P,OPG and RANKL were measured.Results Body weight,Ca,P,RANKL,Tb.Sp of OVX were significantly increased compared to Sham,OPG,BV/TV,Tb.Th,Tb.N decreased.Body weight of H-,M-PSP,Ca and Tb.Sp of PSP,P and RANKL in H-PSP were decreased compared to OVX,OPG in H-,M-PSP,BV/TV,Tb.Th,Tb.N of PSP group increased.The differences were statistically significant (P < 0.05).Conclusion PSP prevents osteoporosis by improving the microstructure of trabecular bone,reducing bone turnover,increasing OPG and reducing RANKL expression.
4.Therapeutic efficacy of small dosage of methylprednisolone in treatment of sepsis and effect on immune cells
Feng XIAO ; yan Zong DANG ; Wan WAN ; ming Zai HE
Drug Evaluation Research 2017;40(9):1331-1333
Objective To investigate the effect of small dosage of methylprednisolone in treatment of sepsis and the influence on immtme cells.Methods Totally 72 patients diagnosed with sepsis in Liqun Hospital from October 2014 to October 2016 were selected in this research.They were randomly divided into two groups:36 patients in control group received conventional anti-infective treatment,and 36 patients in observation group were treated with small dosage of methylprednisolone on the basis of control group.The efficacy and level of immune cells were compared in two groups.Results The survival rate in observation group was significantly higher than that of control group (P < 0.05).The difference of the incidence of complications and the level of immune cells before treatment between two groups had not statistical significance.CD4+ T lymphocytes and CD8+ T lymphocytes in observation group were significantly higher than those of control group after treatment,whereas the ratio ofCD4+/CD8+ and levels of CRP,TNF-α,PCT and IL-1 were lower than those of control group (P < 0.05).Conclusion There is a significant effect of small dosage of methylprednisolone in treatment of sepsis,which can regulate apoptosis of T lymphocyte and inflammatory factor,reduce the immune response,improve the survival rate,and recommend the clinical popularization and application.
5.Molecular characterization of a new mutation E122G of human ornithine transcarbamylase gene.
Hua GAO ; Wei LI ; Zong-he YAN ; Mei-hua JIANG ; De-rong RUI ; Yun-shao HE
Chinese Journal of Medical Genetics 2003;20(1):19-22
OBJECTIVETo determine the molecular basis of late onset ornithine transcarbamylase (OTC) deficiency in a Chinese family of Han nationality and the exon sequences of OTC gene of this patient.
METHODSPolymerase chain reaction-single strand conformation polymorphism and direct sequencing were used to identify the mutation type.
RESULTSA missense mutation E122G in the conserved residue of exon 4 was identified which is unreported before.
CONCLUSIONThe E122G mutation in human OTC gene may cause late onset OTC deficiency.
Age of Onset ; Base Sequence ; Child, Preschool ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Family Health ; Fatal Outcome ; Female ; Humans ; Male ; Models, Molecular ; Mutation, Missense ; Ornithine Carbamoyltransferase ; chemistry ; genetics ; Ornithine Carbamoyltransferase Deficiency Disease ; enzymology ; genetics ; pathology ; Pedigree ; Polymorphism, Single-Stranded Conformational ; Protein Structure, Secondary
6.Regulation of calcium-activated potassium channels of mesenteric artery smooth muscle from patients with essential hypertension by endothelin-1 and prostagl E1.
Xing-lin LUO ; Zong-de WEI ; Chuan HE ; Yan YANG ; Wen-yan WANG ; Xiao-rong ZENG
Chinese Journal of Cardiology 2005;33(1):45-48
OBJECTIVETo study regulation of Ca(2+)-activated K(+) channels (KCa) of mesenteric artery smooth muscle cell (SMC) from 21 old patients with essential hypertension (EH) by endothelin-1 (ET-1) and prostagl E(1) (PGE(1)).
METHODSMesenteric artery branch from EH was digested by enzyme. Patch clamp technique was used to pull cell-attached and inside-out patches on mesenteric artery SMC from EH and the normotensive patients respectively. The signal channel open probability (Po), open dwell-time (To) and close dwell-time (Tc), open channel number per patch were recorded. After adding Ca(2+) (10(-8) approximately 10(-6) mol/L), ET-1(2 approximately 8 x 10(9) mol/L) and PGE(1) (10, 20, 40, 100, 200, 400 nmol/L) to cytoplasm respectively. The parameters above were observed again.
RESULTSCompared to that of normotensive patients, the activities of KCa channels of patients with EH was higher. After adding Ca(2+) to cytoplasm,the Po of KCa channels in normotensive patients increased significantly. But it was few changes in EH group. KCa channels has dual reaction to ET-1 in normotensive patients. We have found no statistics difference when ET-1 present on KCa channels of EH cases. Whereas PGE(1) can affect KCa channels current and channels kinetic significantly in side-out patches. The Po of KCa channels increased. The To protracted and the Tc curtailed in EH.
CONCLUSIONSThe activities of KCa channels of patients with EH increased significantly. but the sensitive to Ca(2+) decreased. ET-1 were few effect to KCa channels. The PGE(1) can activated KCa channels of patients with EH.
Aged ; Alprostadil ; pharmacology ; Cells, Cultured ; Endothelin-1 ; pharmacology ; Female ; Humans ; Hypertension ; metabolism ; physiopathology ; In Vitro Techniques ; Male ; Mesenteric Arteries ; cytology ; Middle Aged ; Muscle, Smooth ; metabolism ; physiopathology ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; drug effects ; metabolism
7.Oxidized high-density lipoprotein promotes maturation and migration of bone marrow derived dendritic cells from C57BL/6J mice.
Zeng-xiang XU ; Yong-zong YANG ; Da-ming FENG ; Shuang WANG ; Ya-ling TANG ; Fan HE ; Yan XIA ; Fang LI
Chinese Medical Sciences Journal 2008;23(4):224-229
OBJECTIVETo explore the influence of oxidized high-density lipoprotein (oxHDL) on the maturation and migration of bone marrow-derived dendritic cells (BMDCs) from C57BL/6J mice.
METHODSThe C57BL/6J mice bone marrow cell suspension was prepared and purified. Recombinant granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant interleukin-4 (rmIL-4) were used to promote monocytes to differentiate and suppress lymphocytes. Then 50 microg/mL oxHDL was added to stimulate BMDCs, using 50 microg/mL high-density lipoprotein (HDL) as homologous protein control, PBS as negative control, and 1 microg/mL lipopolysaccharide (LPS) as positive control. The CD86 and MHCII expression rates were detected with fluorescence-activated cell sorting (FACS). Liquid scintillation counting (LSC) was used in mixed lymphocyte reactions (MLRs) to reflect the ability of BMDCs in stimulating the proliferation of homologous T cells. Levels of cytokines IL-12 and IL-10 were detected by ELISA. The cell migration was evaluated with the transwell system.
RESULTSCompared with PBS group, the expressions of CD86 and MHCII, counts per minute of MLRs, secretion of IL-12 and IL-10, and number of migrated cells in oxHDL group and LPS group significantly increased (all P<0.05), while the increment was less in oxHDL group than LPS group. The number of migrated cells in oxHDL group was about twice of that in HDL group.
CONCLUSIONOxHDL may promote the maturation and migration of BMDCs in vitro.
Animals ; Bone Marrow Cells ; cytology ; drug effects ; physiology ; Cell Differentiation ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; physiology ; Humans ; Lipoproteins, HDL ; metabolism ; pharmacology ; Lipoproteins, LDL ; metabolism ; pharmacology ; Mice ; Mice, Inbred C57BL
8.Illumina sequencing 16S rRNA tagging reveals diverse vaginal microbiomes associated with bacterial vaginosis.
Surong XU ; Lili ZONG ; Mubiao LIU ; Yan HE ; Xuemei HUANG ; Hongwei ZHOU
Journal of Southern Medical University 2013;33(5):672-677
OBJECTIVETo analyze and compare vaginal microbiomes in healthy women at child-bearing ages and patients with bacterial vaginosis (BV).
METHODSA total of 74 vaginal swabs of the vaginal fornix were collected from 37 BV patients and 37 healthy women. BV status was assessed according to Amsels clinical criteria for all the subjects and confirmed using Gram-stain criteria (Nugent scores). Genomic DNA of the samples was extracted for amplifying the 16S rRNA V6 hypervariable region by PCR and pyrosequencing by Illumina. BIPES, UCHIME, TSC and GAST were employed to analyze the information of the species from the samples.
RESULTSLactobacillus was the predominant species in healthy women (more than 95%), including mainly L. iners and L. crispatus, with a small quantity of Gardnerella, Granulicatella, Streptococcus, Prevotella, Escherichia and other genus. The α diversity was significantly increased in 30 BV patients (P<0.001), and β diversity also changed obviously shown by decreased Lactobacillus (varying from 45% to 1%, consisting mainly of L. iners) or even absence Lactobacillus in 6 cases, with increased relative abundance of Gardnerella, Prevotella, Granulicatella, Anaerococcus, Parvimonas, Peptoniphilus.harei, Peptostreptococcus, and Dialister. Different from previous data, 7 BV cases showed a predominance of the rare species L.gasseri and L.acidophilus (75% to 50%).
CONCLUSIONLactobacillus is the predominant vaginal species in healthy women (mainly L. iners and L. crispatus) co-existing with many other bacteria and a variety of microorganisms. Lactobacillus is significantly decreased and even absent in most of BV patients, and some cases show the predominance of the rare species L.gasseri and L.acidophilus.
Adult ; Case-Control Studies ; DNA, Bacterial ; genetics ; Female ; Humans ; Microbiological Techniques ; Microbiota ; Middle Aged ; RNA, Ribosomal, 16S ; genetics ; Reagent Kits, Diagnostic ; Vagina ; microbiology ; Vaginosis, Bacterial ; microbiology ; Young Adult
9.PRMT6 expression in lung of COPD mouse model and its relationship with inflammatory gene expression
Tiao LI ; Qian WANG ; Xue HE ; Ping CHEN ; Shan CAI ; Dandan ZONG ; Yan CHEN
Journal of Chinese Physician 2017;19(11):1611-1614,1617
Objective To explore the expression of protein arginine methyltransferases 6 (PRMT6) in chronic obstructive pulmonary disease (COPD) mouse model and its correlation with inflam mation gene interleukin 6 (IL-6) and cyclooxygenase 2 (COX-2).Methods Sixteen C57BL/8J mice were randomly divided into 2 groups:control group and cigarette smoke extract (CSE)-induced COPD group.Each group was injected intraperitoneally with phosphate-buffered saline solutions (PBS) or CSE at days 1,12,23 and measured lung function and collected lung tissue at day 29.The morphology change of the lung tissue was determined by hematoxylin and eosin (HE) stainning.The protein expression of PRMT6,H3R2me2a and H3K4me3 were detected in lung homogenates by Western-blotting.The mRNA expression of PRMT6,IL-6 and COX-2 were measured by quantitative real-time polymerase chain reaction (qRT-PCR).Results Comparing to control group,COPD group showed typical emphysema changes in the lung tissue,and significantly decreased lung function.The mRNA and protein expression of PRMT6 in the lung tissue of the mice with COPD were significantly decreased,following with the down-regulated signal level of H3R2me2a protein expression,while the increased level of IL-6 and COX-2 mRNA.Meanwhile,PRMT6 was negatively correlated with IL-6 and COX-2 mRNA expression.Conclusions PRMT6 was significantly reduced in CSE-induced COPD mouse model,following with decreased histone H3R2 dimethylation and increased H3K4 trimethylation,negatively correlating with inflammatory gene IL-6 and COX-2 transcription expression.PRMT6 downregulation may activate the transcriptional expression of inflammatory genes involved in the development of COPD,through the regulation of histone methylation level.
10.Mitochondrial damage in early stage of pressure ulcer in rats
Yan ZHOU ; Qing WANG ; Hui HAN ; ze Zong HE ; lan Feng WANG ; mei Feng XING
Basic & Clinical Medicine 2018;38(1):42-46
Objective To investigate the mitochondrial damage and its effect in early stage of pressure ulcer in rats.Methods Forty rats were randomly divided into 5 groups(n=8), control group(Con group) rats without stress, the experimental group was treated with of 170 mmHg for 2 h and relax 0.5 h as one cycle(1C), experi-mental group was divided into 3C, 6C, 9C and 12C group.The pathological changes of the compressed muscle tissue of the rats in each group were observed by HE staining , Western blot was used to detect the expression of Bcl-2 and Bax in the compressed tissue , and the ultrastructure of muscle fibers and mitochondria were observed by transmission electron microscope .Results There were pathological damage and gradually increased in the ex-perimental groups, with the increase of compression cycle; the expression of Bcl-2 in each experimental group was significantly increased as compared with the control group(P<0.05), in the 3C group reached the peak, and then decreased; the expression of Bax was increased gradually with the increase of compression cycle ( P<0.05) , and in the 12C group reached the peak;with the increase of the compression cycle the muscle fibers of each experimental group appeared gradually increased pathological damage:disorder, dissolution and fracture, the ridge of the mitochondria disappeared, vacuolar degeneration, et al.Conclusions In the early stage of pres-sure ulcer in a rat , it brings occurred mitochondrial damage and induces apoptosis .