Objective To detect the encoding gene of efflux pump and two-component system, and investigate the effect of efflux inhibition on the multiresistance of Acinetobacter baumarmii. Methods PCR was used to detect the adeB, adeR and aries gene. Agar dilution was used to determine the minimum inhibitory concentrations(MIC) of ciprofloxacin, cefotaxime, amikacin and imipenem of 50 multiresistance Acinetobacter baumannii, with or without 25 μg/ml reserpine. Results 94%, 96% and 92% of 50 muhiresistance of Acinetobacter baumannii were detected for adeB, adeR and aries gene,respectively. At least four fold decrease of MIC was observed in 49, 50, 50 and 46 isolates for ciprofioxacin, cefotaxime, amikacin and imipenem, respectively. Conclusion The multiresistance of Acinetobacter baumannii is related to the effect of the efflux system.

BACKGROUND: Abnormal lipid metabolism is one of the risk factors in patients with ischemic cerebral disorders, and is correlated with the changes of lecithin cholesterol acyltransferase activity.OBJECTIVE: To observe the relationship between the changes of lecithin cholesterol acyltransferase activity and lipid content in red blood cell membrane.DESIGN: A case-control study(experimental group with control as standard level).SETTING: Department of clinical laboratory, emergency room and department of neurology of a hospital affiliated to a medical college of a university.PARTICIPANTS: Totally 105 inpatients and outpatients with cerebrovascular diseases were selected from the Department of Neurology, Affiliated Hospital of Medical College of Qingdao University, from March 2002 to December 2003. They accorded with the Diagnostic Criteria set at the Second National Conference on Cerebrovascular Diseases. A total of 42 patients with cerebral arteriosclerosis and 63 patients with cerebral infarction were selected as patients group consisting of 67 males and 38 females. Another 65 healthy people receiving physical examination in the hospital, 36 males and 29 females, were selected as control group.METHODS: Venous blood of 8 mL was drawn from the participants on an empty stomach. We assayed the activity of lecithin cholesterol acyltransferase,high density lipoprotein cholesterol, low density lipoprotein cholesterol,apolipoprotein A1 and apolipoprotein B. Red blood cell membrane cholesterol was determined by phthalyl aldehyde-acetometry and red blood cell membrane phospholipid was determined by chemical quantitative analysis.MAIN OUTCOME MEASURES: Changes of lecithin cholesterol acyltransferase activity and lipid content in red blood cell membrane in patients group and control group.RESULTS: According to intention analysis, all the 105 patients in patients group and 65 patients in control group entered the results analysis. Activity of lecithin cholesterol acyltransferase: Activity changes in cerebral arteriosclerosis group and cerebral infarction group were obvious lower than those in control group[(2.14±0.72) kat/L, (2.06±0.80) kat/L, and(2.61± 0. 74) kat/L, P ＜ 0.01 ] . Level of high density lipoprotein cholesterol and apolipoprotein A1: The level in cerebral arteriosclerosis group and cerebral infarction group was obvious lower than that in control group[ (1.32±0.33) mmol/L, (1.37±0.33) g/L, (1.28±0.33) mmol/L; (1.27±0.31) g/L, (1.60±0.43) mmol/L, (1.60±0.43) g/L, t=2.72 to 5.01, P ＜ 0.01 ]. Content of low density lipoprotein cholesterol and red blood cell membrane-cholesterol: The content in cerebral arteriosclerosis group and cerebral infarction group was obvious higher than that in control group [ (2.94 ± 0. 82) mmol/L, (0.63 ±0.05) mmol/g, (3.02 ±0.79) mmol/L;(0.60 ±0.07) mmol/g, (2.56 ±0. 58) mmol/L, (0.57 ±0.05) mmol/g, P ＜ 0. 01 ] . Moreover, the activity of lecithin cholesterol acyltransferase was positively correlated with high density lipoprotein cholesterol and apolipoprotein A1(r=0.247, P ＜0.05; r=0.303, P ＜0.01), but was negatively correlated with low density lipoprotein cholesterol and red blood cell membrane cholesterol(r= -0.212, P ＜0.05;r= -0.346, P ＜0.01).CONCLUSION: In patients with ischemic cerebral disorders, the major change of plasma lipid is the decrease of lecithin cholesterol acyltransferase,but it is not secondary to cerebral infarction. The activity of lecithin cholesterol acyltransferase is positively correlated with high density lipoprotein cholesterol and apolipoprotein A1, but is negatively correlated with low density lipoprotein cholesterol and red blood cell membrane cholesterol.

Objective To evaluate the expression of Sp3 gene of peripheral blood mononuclear cells (PBMC) in multiple sclerosis (MS) patients in Guizhou and the relationship between Sp3 gene expression and immunological function. Methods Two pairs of primers were used to amplify cDNAs generated from 31 MS patients and 30 healthy controls. The serum levels of sIL-2R were measured in 27 patients with MS and 30 healthy controls by sandwiched ELISA. Results The deficient expression of Sp3 gene in MS patients was significantly higher than that in control (41.9% ( 12/31 ) vs 6. 7% (2/30) ,x2 =7. 133 ,P =0. 008). The sIL-2R levels in MS patients were significantly higher than those in control (( 2788.5 ± 1079. 8 ), ( 1270. 7 ± 489. 4) μg/L, t = 6. 170, P = 0. 001 ). The concentration of sIL-2R in MS with negative ((3364.0 ± 1252.3) μg/L) and positive((2450.0 ± 827.0) μg/L) expression of Sp3 gene were significantly increased compared with control (F = 32. 059, P < 0. 05 ). The sIL-2R levels were significantly rising in MS patients with negative expression of Sp3 gene compared with MS patients with positive expression of Sp3 gene ( q = 4. 213, P < 0. 05 ). Conclusions A remarkable deficient expression of Sp3 gene in PBMC has been found in MS patients in Guizhou. sIL-2R may take part in the process of MS. The expression of Sp3 gene is not affected by immune state, however, MS patients with Sp3 deficient expression tend to have a more serious impairment in immunological functions.

Objective To determine the possible genetic background and the source of our hospital's 43 clinical isolates of multidrug-resistant Acinetobacter baumannii, and the category of gene cassettes in type 1 integrons of all strains.Methods Restriction enzyme Apa I was chosed for all strains in pulsed-field gel electrophoresis (PFGE) methods.Multilocus sequence typing ( MLST) was used to compare the allelic profiles of all the strains. PCR method was used for amplify the integrons of all strains. Results PFGE results showed that 43 strains were divided into four types. A-type and B-type were divided into 4 and 2 subtypes, respectively. The MLST results showed the existing of three allelic profiles; 1-3-3-2-2-7-3, 1-3-3-2-2-11-3, and 1-3-3-2-2-14-3.B-type and D-type of PFGE have the same allelic profile(1-3-3-2-2-11-3).A-type strains were detected mainly in ICU, and in burn unit only found B- and D-type.The same integron was detected in 62.8% of the strains.The constituent ratio of A1,A2,A3,A4,B1,B2,C and D-type was 40.7% , 18.5% , 7.4% , 3.7% , 14.8% , 3.7% , 3.7% and 7.4% , respectively.Conclusions The coexistence of multiple cloning system in this region was proved by the PFGE and MLST, and the same clone can evolve to different subtypes when stimulated by different environmental conditions; and the different carrying-situationt of the same integron in strains prove the possibility of the change during the evolution of resistance mechanisms.

OBJECTIVE:To study the in vitro uptake of Resibufogenin(RBG)lactic acid glycolic acid copolymer-water solu-ble vitamin E (PLGA-TPGS) in human liver cancer HepG2 cells,mouse ascites-type lymphatic metastasis of tumor HCa-F cells, and the toxicity on HepG2 cells. METHODS:RCPTN loading RBG and coumarin-6(C6)were prepared. Fluorescent inverted mi-croscope was used to observe the in vitro uptake by RCPTN HepG2,HCa-F cells. It was divided into negative control group,blank PLGA-TPGS nanoparticles(EPTN)group,5-fluorouracil solution(FS)group,RBG solution(RS)group,RBG/PLGA nanoparti-cles(RPN)group and RPTN group. WST-1 was conducted to investigate the optical density at 450 nm wavelength of HepG2 cells after 24,48,72 h incubated by FS,RS,RPN and RPTN with different final concentrations (1.25,2.5,5,10,20 μg/mL);the cell viability (CV) and half inhibitory concentration (IC50) were calculated. RESULTS:RCPTN distributed around the nucleus of HepG2,HCa-F cells. CV was decreased by RBG concentration increased in RPN group and RPTN group,and decreased by time prolonged;compared with FS group,CV in RPTN group was decreased(P<0.05 or P<0.01). IC50 of HepG2 cells incubated by FS,RS,RPN and RPTN was decreased by time prolonged,ordered by RS>FS>RPN>RPTN;IC50 incubated by RPN and RPTN for 48,72 h was obviously less than that of FS and RS(P<0.05 or P<0.01). CONCLUSIONS:RPTN can deliver RBG in-to HepG2,HCa-F cells,showing inhibition effect on HepG2 cells which is stronger than RPN,RS and FS.

Aspirin (AS) has been widely used globally for preventing incidence of cardio-cerebral ischemic disease for nearly 100 years.The people who takes AS for long term may reach several hun-dred million,but many persons were died from interned bleeding.We found salvianolic acids (salvianolic acid B 57％,salvianolic acid A 1％,rosmarinic acid,35％,SA)was much better than AS in preventing in-cidence of cardio-cerebral ischemic disease,and may avoid hemorrhage risk in clinical application.The research are summary briefly as follows: (1) both AS and AS have same anti-platelet aggregation ef-fect,but their mechanism is different.AS inhibited both TXA2 and PGI2,SA inhibited TXA2only;(2)For established thrombosis,SA could dissolved it, AS showed no effect. The thrombolytic mechanism of SA has been elucidated. (3) In SHRSP rats, the incidence of stroke and death rate in SA group was distinct less that of AS group;(4)In MCAO rats,SA and Sal B decreased stroke index and neural im-pairment. AS showed no such ability; (5) There is microcirculatory disturbance in cardio-cerebral isch-emic disease. SA could improve circulatory disturbance induced by LPS, adrenaline, ROS and I/r. there is no any paper reported AS could have beneficial effect on above mentioned microcirculatory dis-turbance models;(6)Hyperlipidemia is an independent risk factor for cardio-cerebral vascular disease. SA could significant hypolipidemic effect which is similar to that of statins(atovastin and simvastin)and ten times stronger than omega-3.AS has no inhibitory effect on hyperlipidemia.(7)Thereis overproduc-tion of ROS induced by ischemic/reperfusion in cardio-cererbal vascular disease.SA showed more ro-bust,anti-oxidant capacity than VitC,Vit E,melatonin,edalavone and resveratrol,etc.SA is one of the most powerful anti-oxidant in the world so far.(8)According to literatures,1/3 patients who take AS for long time will have hemorrhage, we found in normal rats and mice (coagulating and hemodynamics) SA had no apparent effect on coagulation system and this property of SA was confirmed in clinic trial with hundred thousand cases; (9) As well known, neurodegenerative disease are divided into acute and chronic neurodegenerative disease,and both have similar pathogenesis.We proved that SA could inhibit Aβ aggregation and fiber formation, inhibit tau hyperphosphorylation induced by OA and p25/CDK5,as well as increase neurogenesis and angiogenesis.More importantly,SA showed not only pre-ventive effect on cardio-cerebral vascular diseases. SA has been finished clinical trial phase I-IV for treatment of stroke.The therapeutic effect of SA is characterized by inducing multi-target effect and in-hibit pathogenesis of early,middle and late stage of stroke.SA as anti-stroke new drug was approved by the state food and drug administration of China in 2011.

This study was aimed to investigate the effects of Shen-Y uan Y i-Qi Huo-Xue (SYYQHX) capsule com-bined with early percutaneous coronary intervention (PCI) on near-term quality of life (QOL) in unstable angina (UA) patients. Seventy-eight patients diagnosed with UA were selected and randomly divided into the treatment group and control group, with 39 patients in each group. Early PCI was undergone after coronary angiography. Before PCI, the control group was given routine western medication. The treatment group was given routine western medication plus SYYQHX capsule, three pills once, three times daily. The treatment lasted for 30 days. The QOL scores were evalu-ated among patients from both groups before and after treatment in order to determine the efficacy on angina, electro-cardiogram (ECG) and traditional Chinese medicine (TCM) main symptom. The results showed that compared to pre-treatment, the scores of physical limitation (PL), angina stability (AS), angina frequency (AF), and treatment satisfac-tion ( TS ) were significantly increased after treatment ( P < 0 . 05 ) . Compared with the control group , after treat-ment with SYYQHX capsule, the AF, duration time and symptoms of palpitation, fatigue and shortness of breath were obviously improved (P< 0.05). The scores of AS, AF and TS in the treatment group were significantly increased (P < 0.05). It was concluded that SYYQHX capsule combined with early PCI can improve the near-term QOL and TCM main symptoms among UA patients.

In recent decades, scientists in Asian and Western countries have been paying great attention to ginseng research. Today, more than 200 ginsenosides and non-saponin constituents have been isolated and identified. Ginsenosides show biological activities only after being deglycosylated by intestinal bacteria. Aglycone protopanaxadiol and protopanaxatriol show the highest bioactivities. According to literature, the noticeable action of ginseng is that of delaying aging and especially increasing the nootropic effect, and it was found for the first time that Rg1 could increase hippocampal neurogenesis in vitro and in vivo under physiological and pathological circumstances. This is one of primary mechanisms underlying many of its pharmacological actions on the central nervous system. Rg1 was further shown to improve learning and memory in normal rats and mice. The nootropic signaling pathway has also been carried out in normal rats, and the Rg1-induced signaling pathway is similar to the memory formation that occurs in mammals, suggesting that Rg1 may have a potential effect in increasing intellectual capacity in normal people. Comparisons of chemical structures and pharmacologic functions between Panax ginseng and Panax quiquefolium were carried out by many scientists. The conclusion is that each has its own characteristics. There is no superiority or inferiority to the other.
Animals ; Cognition ; drug effects ; Ginsenosides ; pharmacology ; Humans ; Learning ; drug effects ; Memory ; drug effects ; Mice ; Neovascularization, Physiologic ; Neurogenesis ; Neuronal Plasticity ; drug effects ; Panax ; chemistry ; Rats ; Signal Transduction ; drug effects

Objective To investigate the antibiotic resistance and resistance genes of carbapenem-resistant Enterobacteriaceaes (CRE) isolated from 5 hospitals in Northeast China.Methods This study collected 85 CRE isolates during January 2013 to June 2015 from five hospitals in Northeast China.Drug sensitivities of 14 antimicrobial agents were determined by the broth microdilution method.The phenotypes of carbapenemases were screened by modified Hodge test and EDTA test respectively.The genotypes of carbapenemases and other extended spectrum β-lactamases (ESBL) were detected by PCR gene amplification and DNA sequencing method.Using the PCR result as gold standard, the performances of other two carbapenemase detection methods were evaluated.Results Among the 85 CRE strains collected in this study, Klebsiella pneumoniae was the most frequently isolated species (61/85,71.8%).The results of antimicrobial agent sensitivity showed that the 85 CRE strains had resistance rate of cephalosporin and β-lactams/enzyme inhibitor (piperacillin-tazobactam) over 80.0%.The resistance rate of carbapenem was high, with ertapenem 100.0% (85/85), meropenem 65.9% (56/85), imipenem 71.8% (61/85).There were 36 isolates resistant to both meropenem and imipenem.For fluoroquinolones, the resistance rates of levofloxacin and ciprofloxacin were 72.9% (62/85) and 65.9% (56/85), respectively.The resistance rate to fosfomycin and amikacin were 65.0% (55/85) and 54.1% (46/85), respectively.The resistance rate of colistin (21.2%, 18/85) and tigecycline (20.5%, 17/85) were low.Forty-nine strains were modified Hodge test positive and 12 strains were EDTA test positive.By PCR gene amplification and DNA sequencing method, 64 strains carried carbapenemase-encoding genes, of which KPC-2 was the main type (53/85, 62.4%), followed by IMP-4 (10/85, 11.8%), NDM-5 (7/85, 8.2%) and NDM-6 (1/85, 1.2%).At the same time, 85 CRE isolates had the ESBL gene detection and 47 isolates were CTX-M type ESBLs (47/85, 55.3%), with no TEM or SHV type.Conclusions Klebsiella pneumoniae is the majority of CRE strains from 5 large hospitals in Northeastern China.The CRE strains are resistant to most of antimicrobials.Most carbapenemases-producing isolates have the KPC-2 type.Nearly half of the carbapenemase-producing strains also carry ESBL genes, which makes the resistance mechanisms more complicated.

BACKGROUNDTo study the effect of gene amplification and selection system with DHFR plus GS and DHFR or GS gene on the foreign gene expression.

METHODSUsing the N-terminal truncated hTPO(T184) gene as target gene, two plasmidsre were constructed: pDC- T184 and pGC-T184 where DHFR and GS gene were used respectively as the selective amplification marker. They were cotransfected into CHO dhfr cells to establish dual gene amplification and selection system of DHFR plus GS gen and respectively transfected to establish single gene amplification and selection system of DHFR or GS gene. Three selective methods in dual selective system to compare expression efficiency of hTPO were designed: the first method (DG) was to use drug pressure of MTX, then use MSX; the second method (GD) was reversed; the third method was simultaneously to use MTX and MSX as drug pressure.

RESULTSDHFR+GS dual system had not only higher gene amplification efficiency but also higher level expression. There was no distinct affect in different method of drug pressure.

CONCLUSIONSMTX plus MSX dual drug pressure in dual selection system was an efficient and simple method to increase the expression of foreign gene in mammalian cells.

Animals ; CHO Cells ; Cricetinae ; Gene Amplification ; drug effects ; Gene Expression ; Glutamate-Ammonia Ligase ; genetics ; Methotrexate ; pharmacology ; Plasmids ; genetics ; Tetrahydrofolate Dehydrogenase ; genetics