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1.
Zhonghua Yu Fang Yi Xue Za Zhi ; (12): 2122-2128, 2023.
Article in Chinese | WPRIM | ID: wpr-1045987

ABSTRACT

Objective: To explore the drug resistance mechanism and gene structure characteristics of a carbapenemase-producing novel incompatibility group plasmid pNY2385-KPC from Citrobacter freundii. Methods: A multi-drug resistant strain was obtained from urine samples of patients with fever in the emergency ward of Li Huili Hospital, Ningbo Medical Center. Bacterial species was preliminary identified and finally confirmed by 16S rRNA gene amplification and the average nucleotide identity alignment, respectively. The minimum inhibitory concentrations of the antimicrobial agents were determined by VITEK 2 Compact System. The complete genome sequence was obtained by "third-generation" sequencing methods, and then detailed annotation of gene function and comparative genomic analysis of plasmid structure were carried out by BLASTP/BLASTN, RefSeq, ConservedDomains, ResFinder, Isfinder, etc. Results: The pNY2385-KPC carried by citrobacter freundii NY2385 belonged a novel incompatibility group, and contained blaKPC-2 and conjugative transfer (type Ⅳ secretory system, T4SS) genes, which could induce conjugative transfer. A total of 15 plasmids of the same type as pNY2385-KPC were retrieved by NCBI, which were from Citrobacter freundii, and the rest were from Serratia marcescens, Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Raoultella planticola and other bacteria, and were broad-host-range plasmids. The sequence comparative analysis of all 6 of the novel plasmid from Citrobacter freundii showed that the structure of the novel plasmid had certain conserved property, with Tn6296 variant structure carrying blaKPC-2, and plasmid pCF1807-3 had both repApNY2385-KPC and repAIncX8. Conclusion: The pNY2385-KPC type plasmids in Citrobacter freundii carried blaKPC-2 resistance gene, which were divided into two subtypes: repApNY2385-KPC single replicator and repApNY2385-KPC/repAIncX8 complex replicator, belonging to broad-host-range plasmids. And as a mobile genetic element, the plasmids promote the spread of blaKPC-2.


Subject(s)
Humans , Citrobacter freundii/genetics , RNA, Ribosomal, 16S/genetics , Emergency Service, Hospital , Escherichia coli , Genomics
2.
Zhonghua Yu Fang Yi Xue Za Zhi ; (12): 2122-2128, 2023.
Article in Chinese | WPRIM | ID: wpr-1046310

ABSTRACT

Objective: To explore the drug resistance mechanism and gene structure characteristics of a carbapenemase-producing novel incompatibility group plasmid pNY2385-KPC from Citrobacter freundii. Methods: A multi-drug resistant strain was obtained from urine samples of patients with fever in the emergency ward of Li Huili Hospital, Ningbo Medical Center. Bacterial species was preliminary identified and finally confirmed by 16S rRNA gene amplification and the average nucleotide identity alignment, respectively. The minimum inhibitory concentrations of the antimicrobial agents were determined by VITEK 2 Compact System. The complete genome sequence was obtained by "third-generation" sequencing methods, and then detailed annotation of gene function and comparative genomic analysis of plasmid structure were carried out by BLASTP/BLASTN, RefSeq, ConservedDomains, ResFinder, Isfinder, etc. Results: The pNY2385-KPC carried by citrobacter freundii NY2385 belonged a novel incompatibility group, and contained blaKPC-2 and conjugative transfer (type Ⅳ secretory system, T4SS) genes, which could induce conjugative transfer. A total of 15 plasmids of the same type as pNY2385-KPC were retrieved by NCBI, which were from Citrobacter freundii, and the rest were from Serratia marcescens, Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Raoultella planticola and other bacteria, and were broad-host-range plasmids. The sequence comparative analysis of all 6 of the novel plasmid from Citrobacter freundii showed that the structure of the novel plasmid had certain conserved property, with Tn6296 variant structure carrying blaKPC-2, and plasmid pCF1807-3 had both repApNY2385-KPC and repAIncX8. Conclusion: The pNY2385-KPC type plasmids in Citrobacter freundii carried blaKPC-2 resistance gene, which were divided into two subtypes: repApNY2385-KPC single replicator and repApNY2385-KPC/repAIncX8 complex replicator, belonging to broad-host-range plasmids. And as a mobile genetic element, the plasmids promote the spread of blaKPC-2.


Subject(s)
Humans , Citrobacter freundii/genetics , RNA, Ribosomal, 16S/genetics , Emergency Service, Hospital , Escherichia coli , Genomics
3.
Article in English | WPRIM | ID: wpr-787133

ABSTRACT

In vascular smooth muscle, K⁺ channels, such as voltage-gated K⁺ channels (Kv), inward-rectifier K⁺ channels (Kir), and big-conductance Ca²⁺-activated K⁺ channels (BK(Ca)), establish a hyperpolarized membrane potential and counterbalance the depolarizing vasoactive stimuli. Additionally, Kir mediates endothelium-dependent hyperpolarization and the active hyperemia response in various vessels, including the coronary artery. Pulmonary arterial hypertension (PAH) induces right ventricular hypertrophy (RVH), thereby elevating the risk of ischemia and right heart failure. Here, using the whole-cell patch-clamp technique, we compared Kv and Kir current densities (I(Kv) and I(Kir)) in the left (LCSMCs), right (RCSMCs), and septal branches of coronary smooth muscle cells (SCSMCs) from control and monocrotaline (MCT)-induced PAH rats exhibiting RVH. In control rats, (1) I(Kv) was larger in RCSMCs than that in SCSMCs and LCSMCs, (2) I(Kv) inactivation occurred at more negative voltages in SCSMCs than those in RCSMCs and LCSMCs, (3) I(Kir) was smaller in SCSMCs than that in RCSMCs and LCSMCs, and (4) I(BKCa) did not differ between branches. Moreover, in PAH rats, I(Kir) and I(Kv) decreased in SCSMCs, but not in RCSMCs or LCSMCs, and I(BKCa) did not change in any of the branches. These results demonstrated that SCSMC-specific decreases in I(Kv) and I(Kir) occur in an MCT-induced PAH model, thereby offering insights into the potential pathophysiological implications of coronary blood flow regulation in right heart disease. Furthermore, the relatively smaller I(Kir) in SCSMCs suggested a less effective vasodilatory response in the septal region to the moderate increase in extracellular K⁺ concentration under increased activity of the myocardium.


Subject(s)
Animals , Rats , Coronary Vessels , Heart Diseases , Heart Failure , Hyperemia , Hypertension , Hypertrophy, Right Ventricular , Ischemia , Membrane Potentials , Monocrotaline , Muscle, Smooth , Muscle, Smooth, Vascular , Myocardium , Myocytes, Smooth Muscle , Patch-Clamp Techniques , Potassium Channels , Septum of Brain
4.
Article in Chinese | WPRIM | ID: wpr-806087

ABSTRACT

Objective@#To evaluate the application of the central lymph node dissection (CLND) for papillary thyroid carcinoma (PTC) in cN0 T1/T2.@*Methods@#Retrospective analysis of 532 cases with PTC in cN0 T1/T2 who underwent CLND between October 2014 and September 2016 in the Department of Thyroid Surgery, the First Affiliated Hospital of the Kunming Medical University. The incidence of central lymph node (CLN) metastasis and risk factors were analyzed.@*Results@#CLN metastasis rates: 41.2% (42/102) in males vs 34.9% (150/430) in females, P=0.252; 33.9% (116/342) in single focal carcinoma vs 40.4% (74/183) in multifocal carcinoma, P=0.157; 44.0% (125/284) in patients with 45 years old or less vs 27.0% (67/248) in patients more than 45 years old, P=0.000; 30.3% (113/373) in microcarcinoma vs 50.9% (81/159) in non-microcarcinoma, P=0.000.In unilateral lesions, ipsilateral CLN metastasis was correlated with the tumor diameter (P=0.012), but not with the number of lesions (P=0.653). also contralateral CLN metastasis was correlated with the tumor diameter (P=0.000), but not with the number of lesions (P=0.815). For the left or right unilateral single focal lesion, the tumor diameter was not correlated with the metastasis of the posterior to right recurrent laryngeal nerve central lymph nodes (LN-prRLN-CLN) (P=0.652, P=0.088). But in bilateral multifocal carcinoma the tumor diameter was correlated with metastasis of LN-prRLN-CLN (P=0.039).@*Conclusions@#Prophylactic CLND is reasonable for PTC in cN0 T1/T2. A bilateral CLND should be conducted for patients with bilateral multi-focus cancer and unilateral or bilateral non-microcarcinoma, especially in patients more than 45 years old. For unilateral single focal microcarcinoma on the right, the content of CLND should be from laryngeal nerve on right center to posterior branche; for unilateral single focal microcarcinoma on the left side, the left CLND should be conducted. An ipsilateral CLND can be considered in patients with unilateral multifocal microcarcinoma, and generally a routine dissection of the LN-prRLN-CLN is not required, however for bilateral non-microcarcinoma and the the non-microcarcinoma on the right side, the LN-prRLN-CLN dissection should be conducted.

5.
Zhongguo Zhong Yao Za Zhi ; (24): 1189-1193, 2017.
Article in Chinese | WPRIM | ID: wpr-350205

ABSTRACT

To study the inhibitory effect of Rhaponticum uniflorum on apoptosis induced by H2O2 in HepG2 cells. Human HepG2 cells injury models were established by H2O2, then cell survival rate was assayed by MTT method; levels of LDH, ALT, and AST were detected by chemical colorimetric method;SOD activity was detected by xanthine oxidase method; GSH content was detected by dithio-bis-nitrobenzoic acid(DTNB); MDA level was detected by thiobarbituric acid (TBA) method;and the relative activities of Caspase-3, 8 and 9 were measured by Colorimetry. The expression levels of Cleaved Caspase-3(Casp-3), cytochrome(Cyto c), NF-κB, ERK, JNK, p38 MAPK, as well as the phospharylated proteins were determined with Western blotting method. The results showed that R. unifloru had no significant effect on cell viabilities of HepG2 cells at the concentrations of 25-400 mg•L⁻¹. However, H2O2decreased the cell viabilities, increased the cellular oxidative stress, and up-regulated the protein expressions of Casp-3, cytoplasmic Cyto c, p-JNK and nuclear NF-κB. As compared with the model group,R. unifloru could increase the cell viability, reduce LDH, ALT and AST leakage, reduce the MDA formation, increase the SOD and GSH levels,reduce the relative activities of Caspase-3, 8 and 9, down-regulated the protein expressions of Casp-3 and cytoplasmic Cyto c, and down-regulate the p-JNK and nuclear NF-κB levels.The results indicated that R. unifloru had the inhibitory effect on apoptosis induced by H2O2in HepG2 cells, and the mechanism maybe associated with inhibiting JNK activation and NF-κB nuclear translocation.

6.
Article in Chinese | WPRIM | ID: wpr-615994

ABSTRACT

Objective To investigate the value of plasma H-FABP level and Acute Physiology and Chronic Health Evaluation(APACHE Ⅱ) in severity and prognosis estimation for patients with acute pumonary embolism(APE).Methods Totally 160 APE patients were hospitalized from January 2010 to January 2015 and enrolled in this study.According to the severity of the disease,these patients with APE were divided into low-risk group,moderate-risk group and high-risk group.According to clinical prognosis,these patients with APE were divided into survival groups and death groups.Plasma levels of H-FABP were measured by enzyme linked immunosorbent assay(ELISA),and APACHE Ⅱ score were analyzed.The differences of Plasma H-FABP levels and APACHE Ⅱ score were compared and which the relationship with severity and the prognosis of APE were also assessed.Results With the increased severity in patients,the H-FABP and APACHE Ⅱ score were significantly increased (P < 0.05);the H-FABP and APACHE Ⅱ score were significantly higher in death group as compared with survival group(P <0.05).The H-FABP levels and APACHE Ⅱ score were positive correlated(r =0.71,P =0.000).ROC curves analysis results showed that the area under curve of H-FABP was 0.854 (95 % CI:0.784-0.927),and optimal operating point (OOP)was 13.3 μg/L,which had 81.0% sensiticity and 79.4% specificity;ACU of APACHE Ⅱ was 0.861 (95% CI:0.812-0.932),and OOP was 19.2,which had 77.8% sensiticity and 80.4% specificity.The AUC was 0.914 (95% CI:0.825-0.948),and the sensitivity was 88.9%,specificity was 87.6% when the two cutoff values were both achieved,which were higher than the single H-FABP and APACHE Ⅱ score.Conclusion The H-FABP and APACHE Ⅱ score can effectively assess severity and prognosis of APE patients,meanwhile,it provide an objective basis for the clinical individual treatment and reducing the mortality rate of APE patients.

7.
Military Medical Sciences ; (12): 445-448, 2017.
Article in Chinese | WPRIM | ID: wpr-617259

ABSTRACT

Objective To study the transcriptional regulation of vp1667 by H-NS in Vibrio parahaemolyticus.Methods Total RNAs were extracted from Δhns and WT strains.Quantitative RT-PCR was carried out to calculate the transcriptional variation of vp1667 between Δhns and WT.Primer extension assay was also employed to detect the transcription start site and the promoter activity (i.e.the amount of primer extension products) of vp1667 in Δhns and that in WT.The promoter DNA region of vp1667 was amplified, purified, and cloned into the corresponding restriction endonuclease sites of pHRP309 that harbors a gentamicin resistance gene and a promoterless lacZ reporter gene.The recombinant pHRP309 plasmid was transformed into Δhns and WT, respectively, while β-galactosidase activity in cellular extracts was measured using a β-galactosidase enzyme assay system.The over-expressed His-H-NS was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns.The electrophoretic mobility shift assay (EMSA) and DNaseⅠ footprinting were then applied to analyze the DNA-binding activity of His-H-NS to vp1667 promoter region in vitro.Results and Conclusion The primer extension assay detected one transcription start site for vp1667, which was located at 28 bp upstream of vp1667, and its transcribed activity was under the negative control of the H-NS.The EMSA and DNaseⅠ footprinting assay results showed that His-H-NS was unable to bind to the promoter-proximal DNA region of vp1667, suggesting that H-NS indirectly inhibits the transcription of vp1667.

8.
Article in Chinese | WPRIM | ID: wpr-641042

ABSTRACT

Background Cataract with preexisted dry eye is a common eye disease.It is speculated that preservative-free artificial tears can inhibit inflammatory procedure,prevent postoperative eye infections and reduce dry eye symptoms.However,relevant literature is rare up to now.Objective This study was to evaluate and compare the therapeutic effects between preservative-free or preservative sodium hyaluronate combined with fluorometholone eyedrops for cataract with dry-eye.Methods A randomized controlled single-blind clinical study was performed.Sixty patients with dry-eye syndrome who was going to receive surgery for cataract removal were enrolled in Yuhuangding Hospital from January to December 2015 under the informed consent.The patients were randomly divided into the test group and control group.Preservative-free or preservative 0.1% sodium hyaluronate combined with 0.1% fluorometholone eye drops were topically administered in the eyes of the test group and control group,respectively.Ocular Surface Disease Index (OSDI) score,breakup time of tearfilm (BUT),Schirmer Ⅰ test (S I t),corneal fluorescein staining,impression cytology,goblet cell density and the concentrations of interleukin (IL)-1β,tumor necrosis factor (TNF)-α,catalase (CAT),superoxide dismutase 2 (SOD2) in tears were evaluated and compared.Results There were significant differences in gender,ages,OSDI scores,BUT,S I t value,corneal fluorescein staining scores,impression cytology findings,and goblet cell density between the two groups (all at P>0.05).OSDI,corneal fluorescein staining scores and imprint cellular level were evidently reduced,and BUT,S I t values and goblet cell density were significantly increased 1 month and 2 months after operation in comparison with the baseline values in the test group (F =13.058,8.027,3.755,21.652,70.962,92.354,all at P < 0.05).The concentrations of IL-1β and TNF-α in tears of the test group were significantly lower,and CAT and SOD2 in the tears of the test group were significantly higher than those in the control group 1 month and 2 months after operation (F=18.731,9.070,15.357,351.359,all at P>0.05).Conclusions 0.1% preservative-free sodium hyaluronate combined with 0.1% fluorometholone eyedrops can relieve the symptoms and signs of dry-eyes following cataract surgery by playing antiinflammatory and antioxidant effects.

9.
Military Medical Sciences ; (12): 973-977, 2017.
Article in Chinese | WPRIM | ID: wpr-694291

ABSTRACT

Objective To achieve detailed genomic characterization and investigate the antibiotic-resistant mechanisms of plasmid pA1137 carrying the aminoglycoside resistance gene aacC2.Methods Antibiotic-resistant genes were deter-mined by PCR.Conjugation experiments were performed to verify the transferability of plasmid pA 1137.The minimum in-hibitory concentration(MIC)values of bacterial strains were tested with microdilution method.The genetic background, mobile elements and antibiotic resistance mechanisms of pA 1137 were determined using a whole genome sequencing meth-od.Results Both carbapenem-resistant gene blaIMP-8and aminoglycoside-resistant genes aacC2 and aacA4 were carried by A1137 isolated from Enterobacter cloacae(ECL).aacC2 was located in plasmid pA1137 while the other two resistant genes were observed in chromosomes.Plasmid pA1137 was an IncFⅡplasmid,whose total length was 68.97 kb,and GenBank accession number was MF190369.Plasmid pA1137 contained multiple replicons and intact conjugative transfer regions,so it could be transferred into ECL through conjugation experiments and confer corresponding antibiotic resistance to the transconjugant A1137-EC600.Conclusion IncFⅡ plasmid pA1137 has a single accessory region, the first reported Tn5403-based aacC2-tmrB-related region,which can cause stable inheritance and mediate the resistance to aminoglycoside antibiotics in ECL A1137.

10.
Article in English | WPRIM | ID: wpr-727997

ABSTRACT

Ion channels in carcinoma and their roles in cell proliferation are drawing attention. Intracellular Ca2+ ([Ca2+]i)-dependent signaling affects the fate of cancer cells. Here we investigate the role of Ca(2+)-activated K+ channel (SK4) in head and neck squamous cell carcinoma cells (HNSCCs) of different cell lines; SNU-1076, OSC-19 and HN5. Treatment with 1 microM ionomycin induced cell death in all the three cell lines. Whole-cell patch clamp study suggested common expressions of Ca(2+)-activated Cl- channels (Ano-1) and Ca(2+)-activated nonselective cation channels (CAN). 1-EBIO, an activator of SK4, induced outward K+ current (ISK4) in SNU-1076 and OSC-19. In HN5, ISK4 was not observed or negligible. The 1-EBIO-induced current was abolished by TRAM-34, a selective SK4 blocker. Interestingly, the ionomycin-induced cell death was effectively prevented by 1-EBIO in SNU-1076 and OSC-19, and the rescue effect was annihilated by combined TRAM-34. Consistent with the lower level of ISK4, the rescue by 1-EBIO was least effective in HN5. The results newly demonstrate the role of SK4 in the fate of HNSCCs under the Ca2+ overloaded condition. Pharmacological modulation of SK4 might provide an intriguing novel tool for the anti-cancer strategy in HNSCC.


Subject(s)
Carcinoma, Squamous Cell , Cell Death , Cell Line , Cell Proliferation , Head , Ion Channels , Ionomycin , Neck , Neoplasms, Squamous Cell
11.
Military Medical Sciences ; (12): 964-968,973, 2016.
Article in Chinese | WPRIM | ID: wpr-606209

ABSTRACT

Objective To characterize the whole-sequence of plasmid pB557-NDM isolate from Enterbacter cloacae and elaborate its antibiotic-resistant mechanisms .Methods Antibiotic resistance genes were determined by PCR , followed by amplicon sequencing .The activity of class A/B/D carbapenemases was determined by modified Carba NP test .Conjugation experiments were performed to verify the transferability of plasmid pB 557-NDM.The minimum inhibitory concentration (MIC) values of bacterial strains were tested using VITEK 2.The genetic structure, mobile elements and antibiotic-resistant mechanisms of transferable plasmid pB 557-NDM were determined by a whole genome sequencing method .Results The modified CarbaNP test showed that B557 and B557-EC600 had class B carbapenemase activity , and that the blaNDM was carried by plasmid pB557-NDM.This plasmid could be transferred into E.coli through conjugation experiments and therefore could confer corresponding antibiotic resistances to the transconjugant B 557-EC600.Plasmid pB557-NDM was an IncA/C2 plasmid, whose total length was 141.65 kb, and the GenBank accession number was KX786648.It had two inserted regions.One was the blaCMY-6 region where the blaCMY gene was carried by a transposition unit IS Ecp1-blaCMY , the other was the blaNDM-1 region which consisted of a ΔTn1696-In46-rmtC-ISKpn14-ΔTn125 complex structure.Conclusion The production of plasmid pB557-NDM in strain B557 contributes most to its high resistance to many antibiotics .The blaNDM-1 gene is carried in a trancated transposition ΔTn125.

12.
Article in Chinese | WPRIM | ID: wpr-500071

ABSTRACT

Objective To explore the advantages and disadvantages of the double lumen endotracheal intubation and single -lumen endo-tracheal intubation and continuous carbon dioxide insufflation in thoracoscopic esophagectomy .Methods The clinical data of 90 patients in our department of thoracic surgery after thoracoscopic esophagectomy from January 2014 to April 2015 were analyzed .All patients were divid-ed into single-lumen endotracheal intubation (group A)and double lumen endotracheal intubation group (group B).The endotracheal intuba-tion time,operation time,incidence of pulmonary infection,intraoperative and postoperative PaO2,PaCO2,incidence of anastomotic fistula, hospitalization expenses ,length of hospital stay and the incidence of postoperative chylothorax between two groups were compared .Results The difference in intraoperative PaO2,PaCO2,incidence of pulmonary infection,endotracheal intubation time,operation time,hospitalization days and the hospitalization cost between two groups were statistical significance .The difference of the rest index between two groups were no statistical significance.Conclusion Group A has certain advantages in perioperative management ,hospitalization cost and so on,but has disadvantages in perioperative hypoxemia and carbon dioxide retention and acid -base balance disorders .

13.
Military Medical Sciences ; (12): 717-721, 2016.
Article in Chinese | WPRIM | ID: wpr-503986

ABSTRACT

Objective To characterize the resistance mechanisms of a clinical Shigella sonnei strain harboring blaCTX-M-55 .Methods A double-disk synergy test was conducted to detect ESBL.Antibiotic resistance genes were determined by PCR followed by amplicon sequencing.Conjugation experiments were performed to verify the transferability of the plasmids carrying ESBL genes.The minimum inhibitory concentration values were tested using VITEK 2.The transposition unit was confirmed by DNA sequencer,and the transcriptional start site was identified using primer extension assay.Results Strain #1083 produced CTX-M-55,which was encoded by plasmid p1083-CTXM that could be transferred into E.coli through conjugation experiments to confer corresponding antibiotic resistance to the transconjugant #1083-EC600.The transposition unit mediating the transfer of blaCTX-M-55 was ISEcp1-blaCTX-M-55 -Δorf477.ISEcp1 offered strong promoter regions for the resistance genes,facilitating their expressions.Besides,the expressions were constant,not induced by antibiotics.Conclusion BlaCTX-M-55 on plasmids is the major resistance genes for strain #1083.Their expressions and spread are mediated by the insertion sequence ISEcp1.

14.
Article in Chinese | WPRIM | ID: wpr-604847

ABSTRACT

Objective To evaluate the relationship between chemotherapy after pulmonary resection and the incidence rate of bron-chopleural fistula. Methods 246 patients who received pulmonary resection in our hospital from January 2009 to June 2014 were chosen, and they were divided into the chemotherapy group and the non-chemotherapy group. The 138 patients in the chemotherapy group received chemotherapy one month after resection while the other 108 in the non-chemotherapy group did not. Bronchopleural fistula of the two groups were diagnosed and analyzed in order to evaluate the relationship between chemotherapy after pulmonary resection and incidence rate of bron-chopleural fistula. Results There were 12 cases of bronchopleural fistula in the chemotherapy group with an incidence rate of 8. 70%, while there were 2 cases of bronchopleural fistula in the non-chemotherapy group with an incidence rate of 1. 85%. The difference between the two groups is statistically significant (P<0. 05). Conclusion Chemotherapy after pulmonary resection will increase the incidence rate of bron-chopleural fistula.

15.
Military Medical Sciences ; (12): 694-697, 2015.
Article in Chinese | WPRIM | ID: wpr-479179

ABSTRACT

Objective To investigate the regulation of swimming motility by H-NS in Vibrio parahaemolyticus(VP). Methods VP was inoculated into the semi-solid swimming agar plate containing 1% Oxoid tryptone, 2% NaCl, 0.5%Difco Noble Agar, and 0.1% arabinose followed by incubation at 37℃ for 4.5 h before the diameters of bacterial lawns were measured.Total RNAs were extracted from the wild-type (WT) strains and the hns null mutant (Δhns), and the quantitative real-time( RT)-PCR( qRT-PCR) was carried out to calculate the transcriptional variation of flaA between WT andΔhns strains.The entire promoter DNA region of flaA was amplified and cloned into the lacZ fusion vector pHRP309 containing a promoterless lacZ gene. The recombinant lacZ reporter plasmid was transformed into WT and Δhns, respectively, to measure the β-galactosidase activities in cellular extracts using the β-galactosidase enzyme assay system. Results and Conclusion The phenotype results showed that swimming motility of VP was enhanced by H-NS.The qRT-PCR and LacZ fusion results indicated that the transcription of flaA was positively regulated by H-NS.Collectively, H-NS promotes the swimming motility of VP, at least partly, by activating the transcription of flaA.

16.
Military Medical Sciences ; (12): 123-127, 2015.
Article in Chinese | WPRIM | ID: wpr-460195

ABSTRACT

Objective To investigate the carbapenem-resistant mechanism of one strain of Klebsiella pneumoniae( KPN) and one strain of Pseudomonas aeruginosa( PAE) .Methods The identity of the isolates was confirmed by using MALDI-TOF mass spectrometry, 16S rDNA and special gene amplification .The minimum inhibitory concentrations (MICs) of the antimicrobial agents were determined by VITEK 2 Compact System .CarbaNP Antimicrobial susceptibility testing , plasmid extraction, electroporation experiment , PCR amplification, and cloning and sequencing were carried out to analyze the en-coding gene of β-lactamases.Results The types of β-lactamases of the KPN were blaKPC-2 and blaSHV, confirmed by se-quencing of the PCR products , and that of the PAE were blaKPC-2 .Only blaKPC-2 was displayed in both transformants .All of the results of CarbaNP were type A .Conclusion Both strains of KPN and PAE resisting to carbapenem produce a plasmid-mediated carbapenemase blaKPC-2 , which belongs to Bush group 2f, class A β-lactamase.The extended-spectrum β-lacta-mases gene encoding blaSHV of the KPN might be located in the chromosome or not in the plasmid carrying with blaKPC-2 .

17.
Chinese Pharmaceutical Journal ; (24): 469-472, 2014.
Article in Chinese | WPRIM | ID: wpr-859796

ABSTRACT

OBJECTIVE: To investigate the protective effect of different solvent fractions of Boschniakia rossica on acute liver injury induced by acetaminophen (APAP) in mice. METHODS: The mice were randomly assigned to the normal control, model control, bifendate (positive control), as well as groups of different solvent fractions of Boschniakia rossica. Animals were treated once daily for 7d. APAP were given intraperitoneally to the mice of groups, then the alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), reactive oxygen species (ROS), lipid hydroperoxide (LPO), malondialdehyde (MDA), glutathione (GSH), catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), Na+-K+-ATPase, Ca2+-Mg2+-ATPase nitric oxide (NO) and inducible nitric oxide synthase (iNOS) were detected by the colorimetric method. RESULTS: The administration with butanol soluble fraction and aqueous fraction oi Boschniakia rossica reduced the serum ALT and AST activities, increased the scrum ALB level, decreased the hepatic ROS, LPO and MDA levels, increased the CAT, GPx, SOD activities and GSH level, increased the Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of liver mitochondria, and decreased hepatic iNOS activity and NO level of liver in mice with acute liver injury. CONCLUSION: The different solvent fractions of Boschniakia rossica have protective effects on acute liver injury induced by APAP in mice, probably via anti-oxidative and anti-inflammatory activities.

18.
Article in English | WPRIM | ID: wpr-251359

ABSTRACT

This study primarily focused on the systematic assessment of both in vitro and in vivo anti-tumor effects of docetaxel-loaded polyethylene glycol (PEG)2000-polycaprolactone (PCL)2600 micelles on hormone-refractory prostate cancer (HRPC). By using solvent evaporation method, PEG-PCL was chosen to prepare doxetaxel (DTX)-loaded mPEG-PCL micelles (DTX-PMs), with the purpose of eliminating side effects of the commercial formulation (Tween 80) and prolonging the blood circulation time. The prepared DTX-PMs had an average particle size of 25.19±2.36 nm, a zeta potential of 0.64±0.15 mV, a polydispersity index of 0.56±0.03, a drug loading of (8.72±1.05)%, and an encapsulation efficiency of (98.1±8.4)%. In vitro cytotoxicity studies indicated that DTX-PMs could effectively kill LNCap-C4-2B cells and show a dose- and time-dependent efficacy. The hemolysis test showed that DTX-PMs had less hemocytolysis than the commercial product of Duopafei®. A sustained in vitro release behavior and prolonged circulation time in blood vessels were observed in the DTX-PMs. Furthermore, when compared with Duopafei®, the DTX-PMs dramatically reduced the prostate specific antigen (PSA) level and tumor growth of prostate tumor-bearing nude mice in vivo. In conclusion, the DTX-PMs can lower systemic side effects, improve anti-tumor activity with prolonged blood circulation time, and will bring an alternative to patients with HRPC.


Subject(s)
Animals , Humans , Male , Mice , Rats , Antineoplastic Agents , Pharmacokinetics , Pharmacology , Area Under Curve , Cell Line, Tumor , Cell Survival , Dose-Response Relationship, Drug , Guinea Pigs , Hemolysis , Mice, Nude , Micelles , Particle Size , Polyesters , Chemistry , Polyethylene Glycols , Chemistry , Prostatic Neoplasms , Drug Therapy , Pathology , Rats, Sprague-Dawley , Taxoids , Chemistry , Pharmacokinetics , Pharmacology , Treatment Outcome , Tumor Burden , Xenograft Model Antitumor Assays
19.
Article in English | WPRIM | ID: wpr-636512

ABSTRACT

This study primarily focused on the systematic assessment of both in vitro and in vivo anti-tumor effects of docetaxel-loaded polyethylene glycol (PEG)2000-polycaprolactone (PCL)2600 micelles on hormone-refractory prostate cancer (HRPC). By using solvent evaporation method, PEG-PCL was chosen to prepare doxetaxel (DTX)-loaded mPEG-PCL micelles (DTX-PMs), with the purpose of eliminating side effects of the commercial formulation (Tween 80) and prolonging the blood circulation time. The prepared DTX-PMs had an average particle size of 25.19±2.36 nm, a zeta potential of 0.64±0.15 mV, a polydispersity index of 0.56±0.03, a drug loading of (8.72±1.05)%, and an encapsulation efficiency of (98.1±8.4)%. In vitro cytotoxicity studies indicated that DTX-PMs could effectively kill LNCap-C4-2B cells and show a dose- and time-dependent efficacy. The hemolysis test showed that DTX-PMs had less hemocytolysis than the commercial product of Duopafei®. A sustained in vitro release behavior and prolonged circulation time in blood vessels were observed in the DTX-PMs. Furthermore, when compared with Duopafei®, the DTX-PMs dramatically reduced the prostate specific antigen (PSA) level and tumor growth of prostate tumor-bearing nude mice in vivo. In conclusion, the DTX-PMs can lower systemic side effects, improve anti-tumor activity with prolonged blood circulation time, and will bring an alternative to patients with HRPC.

20.
Zhongguo Zhong Yao Za Zhi ; (24): 2187-2190, 2013.
Article in Chinese | WPRIM | ID: wpr-346417

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the protective effect of soyasaponins on acute liver injury induced by D-galactosamine (GalN) and lipopolysaccharide (LPS) in mice.</p><p><b>METHOD</b>The mice were randomly divided into five groups: the normal control, the model group, the silymarin (positive control) group, and soyasaponins high and low-dose groups. They were administered with drugs once every day for 7 days. At the end of the experiment, GalN and LPS were injected intraperitoneally to all of the groups except for the normal group to establish the acute liver injury model. The pathological changes were detected with hematoxylin & eosin (HE) staining, tumor necrosis factor-alpha (TNF-alpha) was detected by ELISA method, and the alanine aminotransferase (ALT), aspartate aminotransferase (AST), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), reduced glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), and the activation of Caspase-3 and Caspase-8 were detected by the colorimetric method.</p><p><b>RESULT</b>Soyasaponins could reduce the activities of serum ALT and AST, the acute hepatic injury induced by GalN/LPS, serum TNF-alpha level, hepatic NO and MDA contents, and the Caspase-3 and Caspase-8 activations of liver tissues, and increase the hepatic CAT, GPx, GST and GSH levels.</p><p><b>CONCLUSION</b>Soyasaponins shows the protective effect on acute liver injury induced by GalN and LPS in mice, which may be related to its antioxidative ability and anti-liver apoptosis.</p>


Subject(s)
Animals , Male , Mice , Alanine Transaminase , Blood , Antioxidants , Metabolism , Apoptosis , Aspartate Aminotransferases , Blood , Caspases , Metabolism , Chemical and Drug Induced Liver Injury , Metabolism , Pathology , Galactosamine , Toxicity , Lipopolysaccharides , Toxicity , Liver , Pathology , Saponins , Pharmacology , Glycine max , Chemistry
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