1.Virulence factors detection and single nucleotide polymorphism assay of extraintestinal pathogenic E.coli in elderly nosocomial infection
Yang CAO ; Shuangqing LIU ; Dianjun WEI ; Wei CHEN
Tianjin Medical Journal 2015;(2):166-169,170
Objective To examine the detection rate of 30 known virulence factors (VFs) of extraintestinal pathogenic Escherichia coli(ExPEC), and to investigates the epidemiology of ExPEC in elderly nosocomial infection. Methods A to?tal of 140 ExPEC clinical isolates from elderly nosocomial patients in hospitals in Tianjin were investigated. Multiplex PCR was performed to detect the 30 virulence factors among the E.coli strains and the detection rate of virulence factors for Ex?PEC were compared between isolates from different sites of infection.Fifty E. coli strains were shown to carry fimH gene that was amplified and sequenced. These sequences were used besides3 references strains (CFT037、UTI89 and K-12 ) to detect SNPs of fimH gene using DNAMAN Version 6.0.3.93 these 53 fimH sequences were used for genotyping and building dendrogram by MEGA4 software. Results In ExPEC, the following virulence factor genes, fimH, traT, fyuA, iutA and kpsMT II, had a higher detection rate than those of the rest . The following virulence factor genes, kpsMT II, K5, papC, pa?pEF ,papG allele II (Internal), papA, cnf1 (CNF), sfa/focDE and rfc had a a higher detectionrate from non-urine origin sam?ples than those from urine origin samples. fimH SNPs analysis of the 50 clinical isolated samples and 3 references samples showed 60 SNPs at 57 polymorphic sites. The fimH SNPs analysis classified the 53 strains into 25 genotype. The genetic fin?gerprintings of 11 isolates were exactly the same. Conclusion Many kinds of virulence factors can be found in ExPEC of el?derly nosocomial infection. The ExPEC strain isolated from non-urine origin had a stronger pathogenicity than those from urine-origin specimens. fimH SNPs analysis is suitable for molecular epidemiological investigation of ExPEC in hospital.
2.Adhesion levels in uropathogenic Escherichia coli with various degree of drug resistance
Yanyan CHEN ; Yang CAO ; Shuangqing LIU ; Wei SONG ; Dianjun WEI
Chinese Journal of Clinical Infectious Diseases 2017;10(1):43-48
Objective To investigate the adhesion levels in uropathogenic Escherichia coli with various degree of drug resistance.Methods One hundred strains of Escherichia coli isolated from urine specimen were collected from patients admitted to 4 Grade A tertiary hospitals in Tianjin during March 2012 to October 2015.Escherichia coli were divided into drug sensitive group and drug resistant group by drug sensitivity tests with 50 strains in each group.The expressions of fimH,fimA,fimB genes of type I fimbriae and papA,papB,papC,papGII genes of P fimbriae were detected by polymerase chain reaction(PCR)and real-time fluorescence quantitative RCR (RT-PCR),respectively.Adhesion ability of type I fimbriae and P fimbriae were tested by yeast cell adhesion test and erythrocyte agglutination test.Chi square test and t(Z) test were used to analyze the data.Results The positive rate of papGII in drug resistant group (42.0%) was significantly higher than that in the drug sensitive group (16.0%)(χ2 =8.208,P <0.05),while there were no significant differences in the positive rates of fimH,fimA,fimB,papA,papB and papC genes between two groups(all P >0.05).The expression levels of fimH,fimB and papC genes in the sensitive group were higher than those in the resistant group(Z =3.427,t =5.182 and 8.120,all P <0.05).The adhesion ability of strains carrying type I fimbriae in sensitive group was stronger than that of resistant group (χ2 =5.769,P <0.05).Conclusions The decline in adhesion ability of type I fimbriae in drug resistant E.coli strains is possibly associated with the adaptive cost of bacteria,the transcription and deficiency of other genes encoded by fim and pap gene cluster will also affect the adhesion function of type I pili and type P pili.
3.Lentiviral vectors with red fluorescent proteins for transfection of human bone marrow mesenchymal stem cells:the red fluorescent protein can express for 21 consecutive days and exert no effects on cell viability
Biao YIN ; Yixi YANG ; Bo YANG ; Le WANG ; Shuangqing LI ; Fuguo YANG
Chinese Journal of Tissue Engineering Research 2015;(10):1539-1543
BACKGROUND:To grasp the optimal multiplicity of infection (MOI) and the time when stronger fluorescence intensities produce can lay the foundation for tracing observation of human bone marrow mesenchymal stem celsin vivo in animal models. OBJECTIVE:To investigate the feasibility of HIV-1 lentivirus carrying enhanced red fluorescent protein to transfect human bone marrow mesenchymal stem cels. METHODS:Human bone marrow mesenchymal stem cels at passage 4 were divided into blank group and MOI 2, 3, 4 groups. After that, the cels were seeded into 12-wel plates at a density of 5.0×105 , and cultured in 1 mL complete medium for adult bone marrow mesenchymal stem cels containing 1% fetal bovine serum. The infectious titer of lentivirus-carried enhanced red fluorescent protein was adjusted to 1.0×10 11 TU/L. Lentivirus solution 10, 15, 20 μL at MOI=2, 3, 4 were respectively added into the MOI 2, 3, 4 groups, and 10μL PBS was added into the blank group. At 24 and 72 hours after transfection, the expression of red fluorescence was observed under an inverted fluorescence microscope and the transfection efficiency was calculated. RESULTS AND CONCLUSION:Enhanced red fluorescent protein expressed stably in bone marrow mesenchymal stem cels. At 24 hours after transfection, red fluorescence could be seen under the inverted fluorescence microscope and achieved the peak at 72 hours after transfection. Within 21 days after transfection, there were no differences in the number of human bone marrow mesenchymal stem cels between the MOI 2, 3, 4 groups and blank group (P > 0.05). These results show that the HIV-1 lentivirus carrying enhanced red fluorescent protein is feasible to transfect human bone marrow mesenchymal stem cels, with the highest transfection efficiency when the MOI=4, which can express at least for 21 days and have no effects on the proliferative activity of labeled cels.
4.Vertebroplasty combined with anti-osteoporosis treatment reduces refracture rate
Fuguo YANG ; Bo YANG ; Biao YIN ; Shuangqing LI ; Yixi YANG ; Yixing GONG
Chinese Journal of Tissue Engineering Research 2016;20(33):4905-4912
BACKGROUND:Currently, vertebral compression fractures are the most common osteoporotic fracture in postmenopausal women;however, incidence of refracture has aroused increasing attention due to a lack of standard treatment. OBJECTIVE:To evaluate whether vertebroplasty combined with anti-osteoporosis treatment can reduce refracture rate fol owing osteoporotic vertebral compressive fractures. METHODS:Eighty-nine patients with osteoporotic vertebral compressive fractures undergoing vertebroplasty were divided into control group (n=38) and treatment group (n=51) after making an informed choice about treatment. Chest/lumbar X-ray and bone mineral density determinations were performed through outpatient or inpatient fol ow-up. The spinal stability, bone mineral density and refracture rate of patients in both groups were fol owed up. RESULTS AND CONCLUSION:Seventy-eight patients achieved complete fol ow-up (ranged from 6-39 months, average 26.73 months). There was no significant difference in the spinal stability between both groups (P>0.05), while rare bone trabecula was found in the control group. There was a significant difference in bone mineral density between both groups at postoperative 12, 24, and 36 months (P<0.05). The refracture rate in the treatment group was significantly lower than that in the control group (P<0.05). Our results indicate that anti-osteoporosis treatment can effectively reduce the incidence of refracture after vertebroplasty in patients with osteoporotic vertebral compressive fractures, and this study found satisfactory short-and medium-term clinical outcomes.
5.Protective effects of metallothionein induced by zinc against doxorubicin-caused cardiotoxicity and its potential mechanisms
Jiabin GUO ; Shuangqing PENG ; Mifeng LIU ; Changhui YAN ; Haiying YANG ; Guoqiang WANG
Chinese Journal of New Drugs and Clinical Remedies 2007;26(2):81-86
AIM: To investigate the effects of metallothionein (MT) induced by zinc on doxorubicin (DOX)-treated mice and to explore the potential mechanisms. METHODS: Male C57BL/6J mice were divided randomly into 4 groups (n = 6) following control, DOX, Zn and Zn plus DOX. Mice were pretreated with eikg-1, ip) or equal volume of saline, and were killed on d 4 after the last injection. Serum and hearts were collected for examination. RESULTS: Zinc pretreatment elevated cardiac MT levels significantly while other antioxidants in heart including glutathione (GSH), glutathione peroxidase (GSHpx) , superoxide dismutase (SOD), and catalase (CAT) were not altered. Severe oxidative injury occurred in the mice treated with DOX as myocardial lipid peroxidation and morphological changes manifested by myocardial fibers swelling and vacuolization and nuclear condensation or dissolution, with increased activities of serum creatine kinase and lactate dehydrogenase and depletion of GSH, GSHpx, and SOD while CAT activity was increased in compensation. However, pre-induction of MT with zinc attenuated all of these toxic changes significantly. Furthermore, DOX induced elevation of hydrogen peroxide in heart tissues was greatly inhibited by zinc pretreatment. CONCLUSION: Preinduction of MT by zinc protects the heart from DOX-induced cardiotoxicity, and this effect is possibly correlated with the property of MT on scavenging free radicals in vivo.
6.T-2 toxin inhibits mitochondrial function of differentiated murine embryonic stem cells
Haiqin FANG ; Lizhong LI ; Zengming ZHAO ; Jun HE ; Jun ZHAO ; Rong YANG ; Xue GENG ; Shuangqing PENG
Chinese Journal of Pharmacology and Toxicology 2014;(3):415-420
OBJECTIVE ToexplorethepossiblemechanismoractiontargetsofT-2toxinembryo toxicity by observing the effect of T-2 toxin on mitochondrial function of differentiated murine e mbryonic stemcells(mESCs).METHODS Duringdifferentiationat24,72and120h,ESCswereexposedto T-2 toxin 0.5 μg·L-1 .Meanwhile,mESCs were pre-treated with antioxidant Trolox (200 μmol·L-1 )for 30 min and exposed to T-2 toxin (0.5 μg·L-1 )for 72 h.The mitochondrial ultrasture of differentiated mESCs was observed under a transi mission electrical microscope (TEM).The differentiated ESC mito-chondrial function,including respiratory control ratio (RCR),ATP synthase activity and mitochondrial membranepotential(MMP),wasmeasuredat144hafterdifferentiation.RESULTS Significant decrease of the mitochondrial number,deformation of mitochondrial structure,and lack of complete mito-chodrial crest were observed through TEM in the groups of T-2 toxin exposed for 72 and 1 20 h,respec-tively.Compared with the normal control group,RCR declined by 49.5% and 55.1%,ATP synthase activity decreased by 84.9% and 89.3%,and MMP decreased by 23.2% and 35.2% in T-2 toxin 0.5 μg·L-1 exposure 72 and 1 20 h group,respectively.However,the inhibition of mitochondrial function by T-2 toxin in differentiated mESCs recovered significantly in the presence of the antioxidant Trolox. CONCLUSION T-2toxininducesoxidativestressandinhibitsmESCsmitochondrialfunctionindifferenti-ated mESCs,and ROS-induced mitochondrial malfunction plays an i mportant role in T-2 toxin e mbryonic toxicity mechanis m.
7.Nephrotoxicity study of Aristolochia fangchi in rats by metabonomics.
Qi LIANG ; Cheng NI ; Ming XIE ; Qi ZHANG ; Yanxia ZHANG ; Xianzhong YAN ; Meijuan YANG ; Shuangqing PENG ; Yuzhong ZHANG
Journal of Integrative Medicine 2009;7(8):746-52
To study the changes of metabolites in rat urine after treatment of Aristolochia fangchi decoction by metabonomic method.
8.Role of gonadotropin-releasing hormone on advancing rat puberty onset induced by estradiol
Chenfeng HUA ; Rong YANG ; Yuxin LLU ; Lina HUANG ; Li ZHANG ; Tianguang ZHANG ; Dongdong TLAN ; Jun ZHAO ; Zengming ZHAO ; Yimei WANG ; Shuangqing PENG
Chinese Journal of Pharmacology and Toxicology 2015;(2):260-264
OBJECTIVE To investigate the role of gonadotropin-releasing hormone (GnRH) in es-tradiol(E2 ) induced advance of puberty onset in rats. METHODS Postnatal day 18 SD rats were given a daily intragastric administration of corn oil or E2(50 μg.kg-1 ) for consecutive 5 d. The day of vaginal opening (VO), pathological changes in ovary and protein expression levels of GnRH, G protein-coupled receptor 54 ( GPR54) and phospholipase C ( PLC) in hypothalamus were observed. RESULTS As compared to corn oil controll group, VO was advanced by about 12.2 d, corpus luteum was observed in the ovary section, and the protein expression levels of GnRH,GPR54 and PLC in hypothalamus were significantly increased by 47%, 55% and 56% in E2 group, respectively. CONCLUSION E2 induced onset of puberty advance may be closely related to regulation of the expression of GnRH, GPR54 and PLC in hypothalamus.
9.Comparative study on metabonomics and on liver and kidney toxicity of Aristolochia fangchi and Stephania tetrandra.
Qi LIANG ; Cheng NI ; Xianzhong YAN ; Ming XIE ; Yanxia ZHANG ; Qi ZHANG ; Meijuan YANG ; Shuangqing PENG ; Yuzhong ZHANG
China Journal of Chinese Materia Medica 2010;35(21):2882-2888
OBJECTIVECombined the blood biochemical markers, the renal histopathological changes and the metabonomics profile were investigated to study the toxicity differences between Aristolochia fangchi and Stephania tetrandra.
METHODTen rats were randomly selected from 70 male Wistar rats as blank control group. The remaining 60 rats were divided into three groups. The two treated groups were orally administrated by 8.1 g x kg(-1) of A. fangchi and S. tetrandra respectively and the control group by equal volume of distilled water for 4weeks. Before the administrated and every 2 weeks, urine and plasma were collected and their 1H-NMR spectra were acquired, and then subjected to data process and PCA. Blood biochemical analysis and histopathological examination were carried out.
RESULTOn the 2nd weekend, the BUN of the two treated groups, the AST of A. fangchi group were all markedly higher than that of the control group (P < 0.05). Compared with the A. fangchi group, the SCr higher in the S. tetrandra group (P < 0.05). The kidney pathological changes were apparently in the two treated groups and the pathological changes in the liver apparently in the S. tetrandra group. Along with the lasting of administration to the 4th week, the BUN, ALT and AST of the two treated groups, the SCr of A. fangchi group were all significantly higher than that of the control group (P < 0.01). The renal and liver injuries in the two treated groups were all become more seriously. Comparing the A. fangchi group, the BUN, SCr and AST were all higher in the S. tetrandra group (P < 0.05). Compared with control group, the urinary concentrations of citrate, 2-oxo-glutarate, taurine, hippurate, TMAO, creatine and the plasma concentrations of 3-D-hydroxybutyrate, acetone, NAC, OAC, creatinine were all changed.
CONCLUSIONThe A. fangchi and S. tetrandra all can induce the renal and liver lesion and its seriousness is correspondent to the lasting of administration. The liver and kidney toxicity of S. tetrandra are all more serious than the A. fangchi.
Animals ; Aristolochia ; chemistry ; Blood Chemical Analysis ; Drug-Related Side Effects and Adverse Reactions ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; metabolism ; Kidney ; chemistry ; drug effects ; metabolism ; pathology ; Liver ; chemistry ; drug effects ; metabolism ; pathology ; Male ; Metabolomics ; Random Allocation ; Rats ; Rats, Wistar ; Stephania tetrandra ; chemistry ; Urine ; chemistry
10.Effectiveness of Longji Daoyin exercise on adolescents of mild idiopathic scoliosis
LIU Lu, ZHANG Wei, HU Zengping, GAO Yang, GU Chuanchuan, YAN Yuntao, DU Shuangqing
Chinese Journal of School Health 2023;44(1):119-122
Objective:
To explore the effectiveness of spinal guidance exercise on adolescents of mild idiopathic scoliosis, so as to provide an experimental reference and method for promoting adolescents spinal health.
Methods:
From the scoliosis screening, 96 children who met the diagnostic criteria were selected in June 2021, and were divided into the experimental group and the control group, with 48 cases in each group. The experimental group received a 12 week Longji Daoyin exercise training, and the control group received health education, observation and monitoring. Before and after the experiment, the axial trunk rotation (ATR) and scoliosis Cobb angle of the two groups were tested respectively, and the subjective sensation of the patients was measured by Scoliosis Research Society-22(SRS-22) questionnaire.
Results:
After 12 weeks, the ATR and Cobb angles of the experimental group decreased [( 5.02± 2.85,7.59±3.94)°],compared to (7.08±2.84)° and (15.84±2.00)° before intervention( t =14.02, 17.09, P < 0.01 ). Scores of functional activity, self image and mental health in the SRS-22 score increased from (4.65±0.14)(3.49±0.21) and (3.99±0.17) to(4.77±0.14)(4.62±0.13)(4.42±0.13)( t =-4.86,-39.12,-13.92, P <0.01 ). The ATR and Cobb angles of the control group increased from (6.94±2.15)° and (15.82±1.76)° to (8.25±2.49)° and (18.63±3.12)°( t =7.44,8.99, P <0.01), and the scores of self image and mental health in the SRS-22 score decreased from (3.49±0.19) and (4.00±0.15) to (3.44±0.18) and (3.94±0.14)( t =-3.74,-4.39, P <0.05).
Conclusion
Early adolescent idiopathic scoliosis is easy to aggravate during the observation period. Longji Daoyin exercise is of preventive and therapeutic significance.