2.Comparison of class 1 integrons detected in Pseudomonas aeruginosa in different periods
Qi-Fa SONG ; Hui LIN ; Jing-Ye XU ; Jian ZHENG ; Chun-Guang JIN ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To study the structure of class 1 integrons in 90 strains of Pseudomonas aeruginosa isolated during two periods of 1992-1996 and 2003-2005,and to get information about the structure changing of class 1 integrons by comparing their structures in two different periods.Methods Routine PCR and long PCR were performed to amplify the class 1 integrons and the gene cassettes they carried, followed with sequencing and blast via GenBank.Results Thirteen out of 41 strians ioslated during the period of 1992-1996 were positive on class 1 intergrons.Long PCR showed that the class 1 integron was 1868 bp in length and contained 2 resistance genes averagely.Six types of resistance genes of qacEA1 (n=6), sull (n=14),aadA1 (n=2),aadB (n=1),PSE-1 (n=2) and tetA (n=1) were found in these integrons, which consisted of 5 patterns of resistance cassette arrangements.Nineteen strains were proved to carry class 1 integrons in 49 isolates from 2003-2005.The mean DNA sequence length of them was 3383 bp with 3.6 resistant genes in averagely,10 types of resistance genes,qacEA1 (n=18),sull (n=25),aadA1 (n=6), aadB (n=7),aacA4 (n=2),PSE-1 (n=3),VEB-1 (n=4),OXA10 (n=1),cm1 A (n=1) and tetA (n =2),were identified in these integrons,which were composed of 9 patterns of resistance cassette arrangements.Conclusion In terms of produce length and resistance cassettes carried in the integrons, greater complexity is found in the structure of class 1 integrons in strains isolated during 2003-2005 than those during 1992-1996.
3.Effects of paroxetine on protein kinase PKA, PKC and CaMKII activity in different brain regions in a rat depression model.
Hui ZHENG ; Guang-yu MA ; Xiao-chun FU ; Hong-guang DU
Journal of Southern Medical University 2008;28(7):1223-1225
OBJECTIVETo evaluate the effects of paroxetine on protein kinase PKA, PKC and CaMKII activities in different brain regions in a rat model of depression.
METHODSThirty-six adult male SD rats were randomized into 6 groups, including one control group (I) and 5 groups of depression model established by forcing the rats to swim for 4 weeks. The 5 depression groups received no treatment (II) or were treated with paroxetine at a single dose (III), for a week (IV), 2 weeks (V) or 4 weeks (VI). The radioactivity of PKA, PKC and CaMKII in the hippocampus and prefrontal cortex was quantitatively measured using a liquid scintillation counter.
RESULTSIn the rat hippocampus, PKA and CaMKII activities were significantly lower in groups II, III, IV, and V than in groups I and VI (P<0.01 or P<0.05), but comparable between groups VI and I (P>0.05). PKC activity was significantly lower in group II than in group I (P<0.01), but showed no significant difference between the paroxetine-treated groups and group I (P>0.05). In the prefrontal cortex, the activity of PKA in groups I, II, III, and IV was similar (P>0.05), but all significantly lower than that in groups V and VI (P<0.01). PKC activity was significantly higher in groups II and III than that in group I and other paroxetine-treated groups (P<0.01), and similar between groups IV and I (P>0.05); groups V and VI had significantly lower PKC activity than group I (P<0.01). Group I had the highest CaMKII activity among the groups (P<0.01).
CONCLUSIONChronic administration of paroxetine can reverse chronic stress-induced inhibition of PKA, PKC and CaMKII activity in rat hippocampus, while the effects of paroxetine on the protein kinases can be more complex in prefrontal cortex.
Animals ; Brain ; drug effects ; enzymology ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; metabolism ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; Depression ; enzymology ; Disease Models, Animal ; Hippocampus ; drug effects ; enzymology ; Male ; Paroxetine ; pharmacology ; Protein Kinase C ; metabolism ; Random Allocation ; Rats
4.Regulatory effect of Chinese herbal compound for detoxifying and activating blood circulation on expression of NF-kappaB and MMP-9 in aorta of apolipoprotein E gene knocked-out mice.
Jing-chun ZHANG ; Ke-ji CHEN ; Guang-juan ZHENG
Chinese Journal of Integrated Traditional and Western Medicine 2007;27(1):40-44
OBJECTIVETo observe the regulatory effect of Chinese herbal compound for detoxifying and activating blood circulation on expression of nuclear factor-kappaB (NF-kappaB) and matrix metalloproteinase-9 (MMP-9) in aorta of apolipoprotein E gene knocked-out (ApoE (-/-)) mice.
METHODSApoE (-/-) mice of 13-week old were divided into two groups and fed with normal diet (Group A) and hyperlipidemic diet (Group B) respectively, the latter was subdivided into 7 groups as Group B1 - 7. Besides, a normal control group was set up with C57BL/6J mice. The drugs used for intervention were polydatin (PD, with 26.6 mg/kg as one dose) for detoxifying and Xiongshao Capsule (XC, with 110 mg/kg as one dose) for activating blood circulation respectively. The intervention was started 19 weeks later by treated Group B1 with PD one dose daily, Group B2 with XC one dose daily, Group B3 with PD and XC each 2 doses daily, Group B4 with PD and XC each one dose daily, Group B5 with PD and XC each half dose daily, Group B6 with lovastatin. To the Group B 7 (as a hyperlipidemia model group) as well as Group A and the normal control group, normal saline was given. After 17 weeks of intervention, the expressions of NF-kappaB and MMP-9 in aorta of mice were determined with immunohistochemical assay.
RESULTSExpressions of NF-kappaB and MMP-9 in aorta and sclerotic plaque were higher in group B7 than those in the normal control group, which were lowered in group B1 - 6 (P < 0.01), and the optimal effect was shown in group B3 (P <0.01).
CONCLUSIONCombined use of Chinese herbal medicine for detoxifying and activating blood circulation could reduce expression of NF-kappaB and MMP-9 in aorta of ApoE (-/-) mice, and the effect of the combination of the two was superior to that of use either of them.
Animals ; Anticholesteremic Agents ; pharmacology ; Aorta ; drug effects ; metabolism ; physiology ; Apolipoproteins E ; genetics ; Blood Flow Velocity ; drug effects ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; Female ; Immunohistochemistry ; Lovastatin ; pharmacology ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Muscle, Smooth, Vascular ; blood supply ; drug effects ; metabolism ; NF-kappa B ; biosynthesis
5.Anatomic assessment of femoral tunnel by transtibial drilling technique in double-bundle anterior cruciate ligament reconstruction: an in vivo study.
Hu XU ; Chun-li ZHANG ; Guang-zheng LI ; Ying-chun WANG ; Hong-bin FAN ; Qin-sheng ZHU
Chinese Journal of Traumatology 2013;16(5):259-264
OBJECTIVETo evaluate the anatomy of femoral tunnels created by simulated transtibial technique in double-bundle anterior cruciate ligament (ACL) reconstruction.
METHODSTwo tibial tunnels, anteromedial (AM) and posterolateral (PL), were drilled 45?and 55?to tibial plateau respectively. On the femoral side, the AM and PL tunnels were drilled through anteriomedial portal. After the four tunnels were established, the shaft of a reamer was introduced into the joint through tibial tunnel and reached against the lateral wall of intercondylar notch. The position that the reamer shaft can reach was marked and recorded.
RESULTSNeither femoral AM nor PL tunnel opening can be fully or partially reached by the reamer shaft through the tibial AM tunnel in all cases. The evaluation through the tibial PL tunnel showed that only in 8 of 50 cases (16%) the femoral AM tunnel opening and in 4 cases (8%) the PL opening can be fully reached. On the other hand, in 12 cases (24%) the femoral AM tunnel opening and in 10 cases (20%) the PL opening can be partially reached by the shafts through the tibial PL tunnel.
CONCLUSIONThe result strongly suggests that transtibial technique is not well competent for femoral tunnel drilling in anatomic double-bundle ACL reconstruction as we have hypothesized.
Adolescent ; Adult ; Anterior Cruciate Ligament ; surgery ; Female ; Femur ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; rehabilitation ; Tibia
6.Expression of choline acetyltransferase in the rat barrel cortex by electrical stimulation.
Hong-Kun FAN ; Chun YANG ; Yan-Yan ZHANG ; Xiao-Ping LE ; Chun-Guang ZHENG ; Li SHI ; Qian ZHANG
Chinese Journal of Applied Physiology 2013;29(4):312-316
OBJECTIVETo observe a turning performance in the rats excited by using a proper electrical stimuli of the barrel cortex region (BC), and the expression of choline acetyltransferase (ChAT) in the BC regions after electoral stimulation.
METHODSSD rats were divided into three groups. The stimulation electrodes were surgically implanted into the bilateral BC regions in the control group and the experimental group rats. The experiment group post surgery for seven days was given the electrical impulses via connection with the electrodes for three times each day through consecutive three days. Three groups of the rats were killed and the brains were quickly removed for frozen sections and then performed with conventional HE and immunohistochemistry staining. And protein samples were prepared from brain and the hippocampus tissues of the three groups to detect the level of the ChAT protein by Western blot.
RESULTSThe experimental rats turn left or right when continuously stimulation in the bilateral BC regions with electric pulse. HE staining showed no significant damage around electrodes in the cerebral cortex. Compared with the control and blank groups, the ChAT positive rate in the brain section in the experimental rats was significantly high by immunohistochemistry assay; the level of the ChAT protein in the rats given the electrical stimulation increased.
CONCLUSIONTurnings performance of the rat could be initiated hy electrical stimuli in the BC region. Expression of ChAT is significantly higher in the BC regions of rat under electrical stimulation, suggesting that acetylcholine might be associated with signal transmission between senses and movement behavior in the nervous central system.
Acetylcholine ; metabolism ; Animals ; Cerebral Cortex ; metabolism ; Choline O-Acetyltransferase ; metabolism ; Electric Stimulation ; Rats ; Rats, Sprague-Dawley
7.Expression of phosphatase of regeneration liver-3 in human colorectal carcinoma and its prognosis value.
Gao-ping ZHAO ; Zong-guang ZHOU ; Wen-zhang LEI ; Cun WANG ; Xue-lian ZHENG ; Yang-chun ZHENG
Chinese Journal of Gastrointestinal Surgery 2008;11(5):487-491
OBJECTIVETo investigate the expression of phosphatase of regeneration liver-3(PRL-3) protein and its relationship with tumor invasion and metastasis in human colorectal carcinoma,and elucidate prognostic value.
METHODSImmunohistochemistry method was applied to detect the PRL-3 expression in the primary tumor specimens and paired paratumor normal tissues from 46 colorectal carcinoma patients, the adenoma tissues from 6 patients with colorectal adenoma, all the metastatic lymph nodes from 29 cases and the metastatic liver lesions from 6 cases. The relationship between PRL-3 expression and clinicopathologic parameters was analyzed and a survival curve was achieved according to Kaplan-Meier method.
RESULTSNo or weak PRL-3 protein expression was detected in normal colorectal mucosa and colorectal adenoma. In colorectal carcinoma tissues, PRL-3 expression was confirmed in 26 of 46 cases (56.5%) of primary colorectal carcinomas (with lymph node metastasis 63.0%, without lymph node metastasis 37.0%, P=0.001), 26 of 29 (89.7%) lymph node metastases, and 5 of 6 liver metastases. The expression of PRL-3 was assembled in the cytoplasm of carcinoma cells and more intensively on the cell membrane.Analysis of the relationship between PRL-3 expression and the clinicopathologic features showed that PRL-3 expression was closely associated with tumor stage (P=0.019), lymph node metastasis (P=0.026), but no relationship with age, sex, tumor size, degree of differentiation was founded (P<0.05). The mean follow-up time was 41.4 months and results showed that patients with positive expression of PRL-3 had a significantly poorer prognosis than those with negative PRL-3 expression group(P=0.032).
CONCLUSIONSPRL-3 protein plays a novel role in tumor progression and metastasis of colorectal carcinoma. PRL-3 can be expected to be a potential predictive biomarker for identifying the prognosis in colorectal carcinoma patients.
Adult ; Aged ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Liver Neoplasms ; metabolism ; secondary ; Liver Regeneration ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; Neoplasm Staging ; Prognosis ; Protein Tyrosine Phosphatases ; metabolism
8.Analysis of beta-lactams-resistance genes in Pseudomonas aeruginosa in burn ward.
Qi-Fa SONG ; Jian ZHENG ; Hui LIN ; Jing-Ye XU ; Chun-Guang JIN ; Guo-Jun LI
Chinese Journal of Burns 2007;23(3):212-215
OBJECTIVETo investigate the resistance genes and antibiotic resistance patterns against beta-lactams in Pseudomonas aeruginosa prevalent in burn ward.
METHODSK-B method was performed to test bacterial resistance patterns against 9 species of beta-lactams in Pseudomonas aeruginosa isolated from wounds and dressings of the patient in burn wards. Seven species of resistance genes against beta-lactams were detected with PCR. Tazobactam-inhibited piperacillin resistance test was performed to study whether the above strains produce extended spectrum beta-lactams.
RESULTSAll 12 strains of bacteria with resistance genes detected were resistant to penicillin and cephalosporins (100%), among them 11 were resistant to all antibiotics. Tazobactam-inhibited piperacillin resistance test demonstrated that all strains with resistance genes were ESBLs.
CONCLUSIONHigh incidence of beta-lactams resistance genes is found in Pseudomonas aeruginosa isolated from burn ward, and they have close relationship with the occurrence of multiple drug-resistance.
Burn Units ; Burns ; microbiology ; Genes, Bacterial ; Humans ; Pseudomonas aeruginosa ; drug effects ; genetics ; isolation & purification ; beta-Lactam Resistance ; genetics
9.Establishment of method for rat hepatocyte primary culture.
Hong-Ling TAN ; Ming-Hui YANG ; Yu-Guang WANG ; Zheng-Chun MA ; Yue GAO
Chinese Journal of Applied Physiology 2006;22(4):509-512
AIMTo explore the culture method of rat hepatocyte between two collagen gel layers on a sandwich configuration and observe the function and morphological characteristics of the hepatocyte, which will be used in evaluation the effect of traditional Chinese Medicine on cytochrome P450.
METHODSRat hepatocyte were isolated by two-step in situ collagenase perfusion method; the hepatocyte were seeded in dishes coated with type I rat tail collagen, culture medium was added and changed daily after gelation. The morphological characteristics of the hepatocyte were observed and biochemical index were tested. The drug effect on the expression of CYP3A was determined by reverse transcriptase-polymerase chain reaction (RT-PCR).
RESULTSRat hepatocytes were successfully attached in gel. BUN and Alb excretion from cell could be tested in the culture period, however, the release of LDH content were lower than the culture system without collagen gel. The typical cellular morphological characteristics of cultured hepatocytes could be observed. PCN increased CYP3A mRNA expression in dose-dependent manner, while the expression of GAPDH wasn't affected.
CONCLUSIONRat hepatocyte sandwich culture can maintenance the cell function and activity, which simulate the environment that more closely in vivo, especially the activity of drug metabolism enzymes.
Animals ; Cell Culture Techniques ; methods ; Cells, Cultured ; Culture Media ; Cytochrome P-450 Enzyme System ; metabolism ; Hepatocytes ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley
10.Toxicity of photoactivated insecticide K-01 to the larvae of Aedes albopictus.
Chun-mei WANG ; Xue-li ZHENG ; Xiao-guang CHEN
Journal of Southern Medical University 2006;26(4):431-434
OBJECTIVETo evaluate the toxicity of photoactivated insecticide K-01 to the larvae of Aedes albopictus and observe the histopathological changes in the larvae.
METHODSThe number of dead larvae was recorded after application of K-01 at different concentrations under different illumination conditions. The content variation of the midgut, malpighian tubules and fat bodies in the larvae was observed microscopically, and the genomic DNA of the larvae was extracted for electrophoresis to identify the target bands.
RESULTSThe maximum larvae-killing effect was achieved with 50 mg/ml K-01 applied under sunlight (100% killing 24 hours after application). Optical microscopic observation of the killed larvae revealed severe damage of the mid-intestinal cells that showed disintegration and elongation. Distinct vacuoles were observed in the fat body cells, in which red droplets were seen to assemble around the cell nuclei. The result of 0.8% agarose gel electrophoresis of the larvae genomic DNA presented typical ladder patterns, suggesting the presence of cell apoptosis.
CONCLUSIONK-01 is an effective photoactivated insecticide.
Aedes ; drug effects ; growth & development ; radiation effects ; Animals ; Insecticides ; toxicity ; Larva ; drug effects ; growth & development ; radiation effects ; Ultraviolet Rays