Objective To analyze the distribution characteristics and antimicrobial resistance of isolated Acinetobacter(A.)bau-mannii from the clinical inpatients in our hospital from 2008 to 2013 to provide the basis for rational use of antibacterial drugs in clinic.Methods The BD phoenix-100 fully automatic microbial analyzer(USA)was adopted to conduct the strain identification and the minimal inhibitory concentration(MIC)detection of clinically isolated A.baumannii.All obtained data were analyzed.Results 128,173,350,282,186 strains of A.baumannii in turn were isolated during these years,the strains in sampling from the clinically i-solated A.baumannii during 2012 was mainly originated from the sputum samples(155 strains,68.28%).In the distribution of hos-pital departments,most concentrated in ICU(65 strains,28.63%),followed by the respiratory department(47 strains,20.70%),the emergency observation wards(35 strains,15.42%)and the apoplexy and geriatric wards(24 strains,10.57%),the most of patients were older people aged more than 60 years.And according to the data within these 5 years,many kinds of different antibacterial drugs had generally the increased drug resistance rate,especially the increasing trend during 2011-2012 was more obvious,the pen-icillins all showed extensive drug-resistance.In the third,fourth generation cephalosporins,aminoglycosides and quinolones were generally drug-resistant.The drug resistance rate of cephalosporins(ceftazidime,cefotaxime,cefepime)remained at a higher level of 75.00%-85.00%.The drug resistance rate of quinolones(ciprofloxacin,levofloxacin)and aminoglycosides(gentamicin,amikacin) also showed the increasing trend year after year.The drug-resistant rate of the beta lactam compound preparations had the rapidly increasing speed,such as which of ampicillin /sulbactam was increased from 60.16% to 83.33%,which of carbapenems such as imipenem and meropenem was increased more significantly,the drug-resistant rate of imipenem was increased from 25.20% to 82.27%,and the detection rate was continually increased.And the drug resistant rate of Schupson was increased from 18.89% to 80%,but the only difference was the drug named Colistin(polymyxin E)maintained a low drug resistance rate during these five years.Conclusion Resistance of A.baumannii to various antibacterial drugs is gradually increased and the isolation rate of multi-drug-resistant and pan-drug resistant A.baumannii is increased year by year.So A.baumannii infection and its drug-resistance changes should be vigilant and be paid more attention to for preventing the emergence and spread of drug-resistant A.baumannii strains.

This study was aimed to explain the relationship between chemical component and therapy objective and to identify effective component group (ECG). The objective of this study is to build a new method to identify the ECG. Traditional Chinese medicine (TCM) formula and one of its treated diseases were selected as the re-search object. The entity grammar systems (EGS) were used as the theoretical framework. A grammatical method for ECG identification was constructed. The component-disease relationship was inferred and the ECG was identi-fied with these inference results. In this paper, 16 compounds which act to 12 proteins in Type 1 diabetes melli-tus (T1D) disease network were identified from Bai-Hu Tang plus Xiao-Ke Fang and 9 chemical compounds were determined as the candidate effective components group. The results indicated that this method can be used to identify ECG and provide a new way to elucidate the molecular mechanism of TCM formula.

Humans ; Estazolam ; Chromatography, High Pressure Liquid

Multi-component traditional Chinese medicines are an innovative research mode for traditional Chinese medicines. Currently, there are many design methods for developing multi-component traditional Chinese medicines, but their common feature is the lack of effective connection of the traditional Chinese medicine theory. In this paper, the authors discussed the multi-component traditional Chinese medicine design methods based on medicinal property combination modes, provided the combination methods with the characteristics of traditional Chinese medicine for the prescription combinations, and proved its feasibly with hypertension cases.
Animals ; Antihypertensive Agents ; administration & dosage ; chemistry ; Blood Pressure ; drug effects ; Drug Combinations ; Drug Therapy ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Humans ; Hypertension ; drug therapy ; physiopathology ; Medicine, Chinese Traditional ; Rats

Background Oxidative damage may cause the functional dysfunction and death of retinal vascular endothelial cells(RVECs),and further leads to the development of retinal vascular diseases.Fufang xueshuantong has a therapeutic effect on retinal vascular diseases,but little is known about its molecular mechanism.Objective The goal of this study was to investigate the protective effects and mechanism of fufang xueshuantong on injury of human RVECs induced by tert-butyl hydroperoxide(t-BHP).Methods Human RVECs were isolated from healthy donor eyes and primarily cultured and then identified by flow cytometry.The third to fifth generations of cells were used in this experiments.The fufang xueshuantong solution of 0.0625,0.1250,0.2500,0.5000 and 1.0000 g/L were added in the cuhure plate with 5 × 104/L cells respectively in the experimental groups,and t-BHP of 75,100,200 and 300 μ.mol/L were added in the model control groups.MTT was used to detect the A490and survival rate of RVECs.The apoptotic rate and death rate of the cells were evaluated by double staining of Annexin V-FITC/PI.Morphology of human RVECs were examined using invert microscopy and Hoechst33258 staining.The expressions of nitro tyrosine (a marker of oxidative damage of protein)and 8-OHdG(a marker of oxidative damage of DNA)in human RVECs were assessed by the immunofluorescence staining.Western blot was used to detect the expressions of nuclear factorkappa B(NF-KB),p53,bcl-2 and bax after 6,12,24 hours t-BHP action.This study was approved by the Ethic Committee of Zhongshan Ophthalmic Center.Results No significant difference was found in A490value among the normal control group,0.0625,0.1250,0.2500,0.5000 and 1.0000 g/L fufang xueshuantong groups(F =1.989,P＞0.05).The survival rates of the cells were lower in 75,100,200 and 300 μmol/L t-BHP groups compared with corresponding fufang xueshuantong groups(t =14.57,13.82,21.51,32.64,P＜ 0.01).The percentages of normal cells were evidently lower in 75,100,200 and 300 μmol/L t-BHP compared with corresponding fufang xueshuantong groups(t=14.908,5.495,17.165,26.330,P＜0.01).The numbers of deformation and death of the human RVECs increased as the elevated concentration of t-BHP,but those in fufang xueshuantong groups were less than the t-BHP groups under the invert microscopy.Compared with t-BHP groups,the expressions of nitro tyrosine,8-OHdG,NF-KB,p53 and bax were lower but the expression of bcl-2 was higher in human RVECs with the statistically significant differences(P＜0.05).Fufang xueshuantong at the concentration of 0.2500 g/L showed maximally protective effect on human RVECs.Conclusions Fufang xueshuantong protects human RVECs against the t-BHP-induced injury through downregulating the expression of NF-kB,p53,bax and up-regulating the express of the bcl-2 protein.

OBJECTIVETo investigate the effect of tetrandrine (TTD) on doxorubicin-induced mdr1 gene expression and its mechanism.

METHODSMTT assay was used to detect the cytotoxicity of TTD to K562 cells. K562 cells were treated with doxorubicin alone or 0.6 microg/ml doxorubicin combined with various concentrations of TTD. RT-PCR was used to detect the mRNA expression of mdr1 and NF-kappa B. Flow cytometry was used to assay the expression of P-glycoprotein (P-gp). Intracellular rhodamine 123 (Rho123) retention assay was applied to test the P-gp function.

RESULTSAfter treatment with 0.6 microg/ml doxorubicin for 24 hours, the expressions of mdr1 mRNA, NF-kappa B mRNA and P-gp in K562 cells were increased from 0.171 +/- 0.012, 0.783 +/- 0.090, 7.85 +/- 0.15 to 0.428 +/- 0.012, 1.075 +/- 0.047 and 73.68 +/- 1.84, respectively. The intracellular Rho123 retention was decreased from 711.9 +/- 63.6 to 347.8 +/- 60.6, indicating up-regulation of P-gp function (P<0.05). Pretreatment of K562 cells with 2.0 microg/ml TTD for 24 hours and then incubated for another 24 h with doxorubicin, the expressions of mdr1 mRNA, NF-kappa B mRNA, P-gp and up-regulation of P-gp function induced by doxorubicin were prevented in K562 cells (0.148 +/- 0.006, 0.627 +/- 0.098, 7.18 +/- 0.38 and 799.7 +/- 45.8, respectively P<0.05). But 0.5 microg/ml and 1.0 microg/ml TTD had little effect.

CONCLUSIONSTTD inhibits the expression of mdr1 mRNA, P-gp and up-regulated P-gp function induced by doxorubicin in a dose dependent manner. The mechanism of this effect may be down-regulation of NF-kappa B by TTD.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Benzylisoquinolines ; pharmacology ; Doxorubicin ; pharmacology ; Humans ; K562 Cells ; NF-kappa B ; metabolism ; RNA, Messenger ; genetics ; Up-Regulation ; drug effects

Objective To explore the effect of dendritic cells (DC) modified with transforming growth factor β1 (TGF-β1) gene on the expressions of CD28/CTLA-4:B7 costimulatory molecules in peripheral blood mononuclear cells (PBMC) in the Lewis rats with experimental autoimmune myasthenia gravis (EAMG). Methods Thirty inbreeding line, healthy, female Lewis rats were divided randomly into 6 groups: normal group, EAMG group, DC treatment group, pcDNA3-TGF-β1-DCtreatment group, pcDNA3-DC control group and normal saline group. The rats were immunized with the AChR protein extracted from electric organ of Narcine timilei and CFA in the groups except normal group. 2×106 pcDNA3-TGF-β1-DCs/rat were injected subcutaneously into the backs of the rats which had been immunized 5 d earlier with AChR+CFA. The rats in DC treatment group, pcDNA3-DC control group and normal saline group were injected in parallel with untreated DC, pcDNA3-DC and normal saline, respectively. Seven weeks after the first immunization, the expressions of CD28 mRNA and CTLA-4 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the levels of B7-1 and B7-2 on the surface of PBMC were examined using flow cytometry. Results (1)The low expression of CD28 mRNA and rare expression of CTLA-4 mRNA were found in the normal rats, and both expressions increased markedly in EAMG rats (P＜0.001). Compared to those in EAMG group, the expression of CD28 mRNA decreased and CTLA-4 mRNA was upregulated after the treatment with pcDNA3-TGF-β1-DC (P＜0.05). There was no significant difference in the expressions of CD28 mRNA and CTLA-4 mRNA among the EAMG group, DC treatment group, pcDNA3-DC control group and normal saline group (P＞0.05). (2) The expressions of CD28, CTLA-4, B7-1 and B7-2 on the surface of PBMC were rare in normal rats, which increased significantly in EAMG rats (P＜0.001). The levels of CD28, B7-1 and B7-2 in pcDNA3-TGF-β1-DC group were lower than those in EAMG group (P＜0.01), but the level of CTLA-4 was higher than that in EAMG group (P＜0.05). They showed no statistically difference among the EAMG group, DC treatment group, pcDNA3-DC control group and normal saline group (P＞0.05). Conclusions The expressions of CD28/CTLA-4:B7 costimulatory molecules are abnormal in the rats with EAMG. The regulation of CD28/CTLA-4:B7 costimulatory pathways may play a critical role in the mechanism of the treatment with DC transfected with pcDNA3-TGF-β1 in the incipient EAMG rats.

Objective To investigate the differences of blood glycosylated hemoglobin (HbA1c) levels between the patients with acute cerebral infarction and healthy controls, and explore the relation between HbA1c level and both neurological deficits scores and imaging of lesions in number. Methods One hundred and eighty-six patients with acute cerebral infarction within 1 week were performed neurological deficits scales after the admission; the HbA1c level was measured within 24 h admission; brain MRI scan was performed on these patients 48 h after onset or stabilization. Glucose tolerance test was taken at the rehabilitation of infarction (except for having a clear history of diabetes before). At the same time, 160 healthy controls were checked on the level of HbA1c and taken the glucose tolerance test. The differences of blood HbA1c levels between the patients with acute cerebral infarction and healthy controls were investigated; and the relation between HbA1c level and both neurological deficits scores and imaging of lesions in number was explored. Results The HbA1c level in patients with acute cerebral infarction (6.982%±1.803%) was significantly higher than that in the controls (5.128%±0.592%, P<0.05). The level of HbA1c in patients with cerebral infarction and the neurological deficits scores were positively correlated (r=0.760, P<0.05). The level of HbA1c in patients with 2 lesions (6.635%±0.427%) was obviously higher than that in patients with 1 lesion (5.803%±0.307%, P<0.05); The level of HbA1c in patients with 3 or more lesions (8.571%±0.519%) was obviously higher than that in patients with 1 or 2 lesions (P<0.05). Conclusion Diabetes is a major risk factor for cerebral infarction. High HbA1c level might cause a series of cerebrovascular diseases, thus it is an important factor in the happening of cerebral infarction and HbA1c level is an important indicator of the early assessment of the severity of the diseases. The incidence of cerebral infarction can be decreased by controlling hyperglycaemia, lowering the HbA1c levels.

This study was purposed to explore the mechanisms of preventive effect of tetrandrine (TTD) on doxorubicin (ADM)-induced multidrug resistance (MDR) in human leukemia cell line K562 from two aspects of the transcription control of MDR1 gene and cell apoptosis. The experiment was divided into 3 groups: group I-blank control; group II-ADM-induced drug-resistance; group III-ADM-induced drug-resistance after pretreatment with TTD. Reverse transcription-PCR (RT-PCR) was used to detect the mRNA expression levels of c-Jun, YB-1 and Survivin genes. Western blot was used to determine the nuclear protein expression levels of c-Jun and YB-1. Flow cytometry was used to assay the apoptosis of cells. The results showed that as compared with group I, the expression levels of c-Jun mRNA and nuclear protein decreased (p < 0.05), as well as the expression levels of YB-1 mRNA and nuclear protein increased in group II (p < 0.05). However, the expression of Survivin mRNA had no change (p > 0.05); the apoptosis rate of cells was 8.31%. As compared with group II, the expression levels of c-Jun mRNA and nuclear protein increased (p < 0.05), expression levels of YB-1 mRNA and nuclear protein as well as Survivin mRNA decreased in group III (p < 0.05). The apoptosis of cells was 97.2%. It is concluded that TTD can inhibit the expression of YB-1 and up-regulate the expression of c-Jun, thus inhibit the expression of MDR1 gene. TTD can also inhibit the expression of Survivin and increase the apoptosis of cells induced by ADM.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Apoptosis ; drug effects ; genetics ; Benzylisoquinolines ; pharmacology ; Drug Resistance, Multiple ; drug effects ; genetics ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; K562 Cells ; Proto-Oncogene Proteins c-jun ; metabolism ; Y-Box-Binding Protein 1 ; metabolism

Experiments were performed in male Sprague-Dawley rats (150-200 g). A silver clip (0.2 mm internal diameter) was placed on the left renal artery of rats. After operation the rats were divided into 4 groups sham group, 2K1C (two-kidney one clip) group, 2K1C+Arg (2K1C and L-Arg 150 mg/kg x d(-1) by drinking) group, and 2K1C+NAME (2K1C and L-NAME 10 mg/kg x d(-1) i.p.) group. The animals were studied at two time points (4 and 7 weeks after operation) corresponding to phases I and II of Goldblatt hypertension. The animals were deeply anesthetized with pentobarbital and perfused by the cardiac route with saline (100 ml) and freshly prepared 4% paraformaldehyde in phosphate buffer (300 ml). The caudal medulla was removed, then placed in 25% sucrose in PB for 12 h in a 4 degrees C fridge. The 40 microm coronal slices of brainstems were cut with cryostat, collected in PBS for nNOS study by immunohistochemistry. The results showed that (1) only a few neuronal nitric oxide synthase (nNOS) positive neurones were found in caudal medulla, including the depressor area of the ventral surface of medulla oblongata (VSMd) and the caudal pressor area (CPA) of the sham operated animals. The number of nNOS positive neurons in caudal medulla was significantly increased in 2K1C Goldblatt hypertension rats at 4 and 7 weeks. (2) The number of nNOS positive neurons in VSMd and CPA were 2K1C+Arg > 2K1C >2K1C +NAME > sham. (3) L-Arg enhanced the expression of nNOS whereas L-NAME inhibited the expression of nNOS in caudal medulla. (4) The character of nNOS expression was similar in Goldblatt hypertension rats at 4 weeks with that of the rats at 7 weeks.
Animals ; Hypertension, Renovascular ; enzymology ; physiopathology ; Male ; Medulla Oblongata ; enzymology ; metabolism ; Nitric Oxide Synthase ; biosynthesis ; Rats ; Rats, Sprague-Dawley