Bacterial Infections ; epidemiology ; microbiology ; China ; Drug Resistance, Bacterial ; Humans ; Microbial Sensitivity Tests

Objective To investigate the effect of MDR1 and MDR3 gene silence by shRNA of human breast carcinoma cell line MCF-7/Adr,and explore the role of MDR1 and MDR3 in adriamycin-resistance of breast carcinoma cells. Methods shRNA plasmid vector specifically targeting MDR1 and MDR3 gene was transfected into cells. The control group was transfected with empty vector. The concentration of adriamycin was detected by the flow cytometry (FCM). Cell apoptosis was analysed by FITC-Annexin-V/PI double staining. Cell viability and the IC50 of adriamycin on MCF-7/Adr cells were determined by MTT method. MDR1 and MDR3 mRNA were assessed by RT-PCR. P-gp expression was detectedby immunochemistry. Results After treatment with ABCB1 and ABCB4 shRNA plasmid vector, the apoptosis of MCF-7/Adr cells was (30.21±1.65)%and (22.07±2.17)% respectively. Compared with untransfecedgroup and empty vector transfection group the difference was significant(P＜0.01). MDR1 and MDR3 shRNAcould increase cellular adriamycin accumulation of MCF-7/Adr cells. MCF-7/Adr cells viability and the IC50were significantly decreased after transfection. Compared with untransfeced group and empty vector transfectiongroup, the mRNA level of MDR1 and MDR3 in MCF-7/Adr cells were decreased by (89.5±0.8)%and(85.1±1.2)%, the reduction of MDR1 and MDR3 mRNA was in a time-dependent manner. Immunochemistry proved that the expression of p-gp was significantly inhibited. Compared with untransfeced group and empty vector transfection group the difference was significant (P＜0.05). Conclusion The shRNA can effectively and specifically silence the expression of MDR1 and MDR3 gene, reverse the adriamycin-resistance mediated by P-gp in MCF-7/Adr cells. The reversal effect of adriamycin-resistance by shRNA of MDR1 is more effective than that of MDR3.

Objective To study the distribution of variable number tande repeat(VNTR) polymorphisms of the platelet membrane glycoprotein Ⅰ b? in Han nationality at Harbin and the relationship between these polymorpbisms and cerebral infarction(CI).Methods The identification of alleles and genotypes of VNTR polymorphism of the glycoprotein Ⅰ b? gene was performed by polymerase chain reaction (PCR)in 200 healthy individuls and 200 CI patients(77 lacunar infarction patients and 123 atherosclerotic thrombotic infarction patients),to analyze The relationship between gene polymorphisms and cerebral infarction.Results(1)There were three types of alleles:B、C、D,and five types genotypes:BC,BD,CC, CD,DD in Harbin Han nationality.No person with A allele and BB genotype was found.(2)No statistically significant differences of GP Ⅰ b? gene VNTR polymorphism was found between CI patients or subtype CI patients and controls(P=0.412 and 0.572,respectively).Conclusions(1)This study indicates that the C and D alleles of VNTR polymorphisms of GP Ⅰ b? are the main alleles while the CC and CD genotypes are the main genotypes in Harbin Han people.(2)Our findings indicate that no association exists between the VNTR polymorphism of platelet GP Ⅰ b? gene and CI.

OBJECTIVETo investigate the relationship between activities of early growth response gene 1 (EGR-1) of p38 mitogen-activated protein kinase (MAPK) pathway and in the epirubicin resistance of breast carcinoma cells.

METHODSProtein expression of phosphorylated p38MAPK was detected by confocal spectral microscopy. Using specific inhibitor SB203580, the effect of p38MAPK on cell apoptosis was analyzed by FITC-Annexin-V/PI double staining. The concentration of epirubicin was detected by flow cytometry (FCM). The 50% inhibition concentration (IC50) of epirubicin on MCF-7/Adr cells was determined by MTT method. Electrophoretic motility shift assay (EMSA) was performed to examine the affinity of EGR-1. EGR-1 mRNA was assessed by RT-PCR. The expression levels of p-glycoprotein, phosphorylated p53 and p38 were detected by Western blot.

RESULTSAfter treatment with SB203580 (15 micromol/L) 24 h and 48 h, (1) the early and late apoptosis of MCF-7/Adr cells expressing the phosphorylated p38MAPK protein was (25.36 +/- 1.17)% and (38.21 +/- 1.25)%, respectively, P < 0.05. And the tendency was in a time-dependent manner. (2) The average fluorescence intensity of MCF-7/Adr cells expressing the phosphorylated p38MAPK protein was (32.45 +/- 2.36) and (41.66 +/- 3.12), higher than the blank group (14.17 +/- 1.45) and DMSO group (16.28 +/- 0.63), P < 0.01. The epirubicin resistance of MCF-7/Adr cells significantly decreased. (3) SB203580 demonstrated a significantly higher level of EGR-1 activity. The IC50 was (21.53 +/- 2.17) and (8.77 +/- 1.02), lower than the DMSO group (40.74 +/- 2.56). MCF-7/Adr cells treated with SB203580 down-regulated the p38MAPK pathway activity, but up-regulated the EGR-1 mRNA expression. SB203580 significantly increased the cellular phosphorylated p53 protein level, but decreased the p-glycoprotein level in MCF-7/Adr cells.

CONCLUSIONSThere is a close relationship between p38MAPK pathway activity and the epirubicin resistance of breast carcinoma cells. The activation of EGR-1 mediated by p38MAPK pathway plays a critical role in epirubicin resistance.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Apoptosis ; drug effects ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Early Growth Response Protein 1 ; genetics ; metabolism ; Enzyme Inhibitors ; pharmacology ; Epirubicin ; pharmacology ; Female ; Humans ; Imidazoles ; pharmacology ; Phosphorylation ; Pyridines ; pharmacology ; RNA, Messenger ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

This paper proposes a one-way micro valve with a simple structure and a simulation design for the engineering capsule. We have now got its design parameter selection method and its mechanic characteristic from experiments.
Capsules ; Computer Simulation ; Drug Delivery Systems ; methods ; Equipment Design ; Infusion Pumps, Implantable ; Technology, Pharmaceutical ; instrumentation ; methods

OBJECTIVETo observe the effect of β₃adrenoceptors (β₃-AR) activation on rat thoracic aorta smooth muscle contractility and the possible related mechanism.

METHODSThe endothelium removed thoracic aorta was pre-contracted with 30 mmol/L KCl physiological saline solution (PSS). Then the tension of the thoracic aorta was recorded in presence of BRL37344 (BRL) to determine the action of β₃-AR. The tension of the thoracic aorta was also recorded in the presence of Propranolol (PRA), SR59230A (SR), L-NNA, H-89 and Iberiotoxin (IBTX) respectively to reveal the underling mechanism of β₃-AR activation on rat vascular smooth muscle. Immunohistochemistry was adopted to confirm the existence and the distribution of β₃-AR in rat thoracic aorta.

RESULTSThe results showed that: (1) The thoracic aorta was relaxed by β₃-AR activation, with a relaxation percentage of (10.59 ± 0.79). (2) β₃-AR was expressed in both endothelial and smooth muscle layer in thoracic aorta sections of rats. (3) PRA did not block the effect of BRL on the thoracic aorta. The relaxation actions of BRL could be antagonized by pre-incubating the thoracic aorta with SR. (4) L-NNA (a NOS inhibitor) and H-89 (a PKA inhibitor) reversed the relaxation effect of BRL on vascular smooth muscle. (5) The effect of BRL was decreased after application of Ibriotoxin (IBTX), a large conductance calcium dependent potassium channel blocker.

CONCLUSIONThe results confirmed that activation of β₃-AR led to relaxation of thoracic aorta smooth muscle. The relaxation action of β₃-AR on smooth muscle of rat thoracic aorta was related to activation of NOS and PKA signaling pathway. Large conductance Ca²⁺-K⁺ channels were involved in the relaxation action of β₃-AR activation on rat thoracic aorta smooth muscle.

Animals ; Aorta, Thoracic ; physiology ; In Vitro Techniques ; Isoquinolines ; Large-Conductance Calcium-Activated Potassium Channels ; physiology ; Muscle Contraction ; Muscle Relaxation ; Muscle, Smooth, Vascular ; physiology ; Nitroarginine ; Peptides ; Propanolamines ; Propranolol ; Rats ; Receptors, Adrenergic, beta-3 ; physiology ; Signal Transduction ; Sulfonamides

OBJECTIVETo investigate the effect and mechanism of Scutellaria Baicalensis Stem-leaf Total Flavonoid (SSTF) on lipid metabolism in hyperlipidemic rat.

METHODOn the basis of establishing hyperlipidemia rat model, blood lipids, lipid metabolic enzyme, antioxidative capacity were investigated after 30 days feeding of fatty emulsion.

RESULTSSTF significantly reduced the serum TC, TG, LDL-C, ApoB concentration and increased HDL-C and ApoAI levels, improved the activity of lecithin cholesterol acyltransferase (LCAT). SSTF was shown to decreased MDA content in both serum and liver, increased serum SOD activity.

CONCLUSIONSSTF could remarkably modulate the lipid metabolic disorder in hyperlipidemic rats, and has a certain regulating function on lipoprotein, inferring that it could reduce the occuring of atherosclerosis. The mechanism of regulating lipid metabolism might be related with the increasing activity of LCAT and antioxidative capacity.

Animals ; Apolipoprotein A-I ; blood ; Apolipoproteins B ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Flavonoids ; chemistry ; pharmacology ; Lipid Metabolism ; drug effects ; Male ; Plant Extracts ; chemistry ; pharmacology ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Rats ; Rats, Sprague-Dawley ; Scutellaria baicalensis ; chemistry ; Triglycerides ; blood

Objective To evaluate the clinical efficacy of standardized house dust mite allergen specific immunotherapy (SIT) for rhinitis and asthma in children. Methods Forty-two children with allergic rhinitis and asthma who received a standardized house dust mite allergen SIT in our hospital were enrolled in our study. The result of allergen skin prick test, serum specific IgE levels (sIgE) of house dust mite and dust mite,pulmonary function and symptom scores were analysed before and at one year after treatment in all children. Results Skin indexes of house dust mite and dust mite, symptom score were significantly decreased at one year after treatment,but the levels of house dust mite and dust mite sIgE,lung function test (FVC,FEVt,MEF25-75) showed no significant differences. Conclusion Children with allergic rhinitis and asthma have significant improvements in their skin sensitivity and clinical symptoms by given SIT for one year,but the impact of SIT on airway inflammation needs further observation.

Objective To investigate the thermal effects on tissue structures during microwave endometrial ablation(MEA)and seek a feasible method of endometrial thinning and a fitting mode of applicator radiating microwaves.Methods Operations were performed between the group of thorough uterine curettage and the group of early follicular phase in in vitro or in vivo uterus.The former was treated with MEA after thorough uterine curettage;while the latter was treated with MEA in the early follicular phase directly.The applicator radiating microwaves were moved in "Z" or "Z+W" shape inside uterine cavity.At the same time the serosal temperature was measured in the uterine fundus,tael cornua uteri,the posterior wall and the lower part of anterior wall.After operations the uterine specimens were stained by hematoxylin- eosin,and respiratory enzyme dehydronicotinamide adenine dinucleotide phosphate diaphorase(NADH-d) methods.The morphologic changes and the depth of tissue thermal damage were evaluated using an optical microscope and electron microscope.Results(1)Under the optical microscope the endometrial glands became distorted,the cell boundaries disappeared,the nucleoli turned condensed and were stained darker. A large number of acute inflammatory cells appeared in fibrous tissue.In the shallow muscle layer cells were arrayed thickly,nucleoli were solidified and condensed,and cellular plasm were concentrated.The endometrial and the superficial muscle layers were damaged and colorless with NADH-d staining.The scope of the tissue thermal damage was clearly seen.Under an electron microscope,some smooth muscle ceils, chromatin,karyotheca and cellular membranes were destroyed.The mitochondria were swollen,membranes were ruptured,and the crista disappeared.Many organelles were destroyed.The chromatin was lightly wrecked in the transitional area between putrescence and the normal smooth muscle tissue.Karyotheca and cellular plasm still existed,the mitochondria were highly edematous and the crista were disappeared,and the granular endoplasmic reticula were slightly expanded and degranulated.(2)The serosal temperature in in vitro uterus was significantly higher than that in in vivo uterus(P0.05).The injury depth of the "Z+ W" radiation group increased significantly than that of the " Z" radiation group(P

The cloning of promoter is important for studying the genetic engineering and the regulation of gene expression in plants. Two promoters Os772 and Os359, which are predicted to be highly expressed in the endosperm of rice from the EST database were cloned. After construction of the Os772∶∶GUS and Os359∶∶GUS expression vectors, they were transformed into rice. X-Gluc staining of transgenic plants showed that Os772 and Os359 can promote GUS gene expression in matured endosperm but not in root, stem, leaf and flower. This result indicates Os772 and Os359 are two rice endosperm-specific promoters.