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 pharmacokinetic features of silibinin from Jiqi Injection(JI) in Beagle dogs,and to observe the effect of Panax notoginseng saponins,another active component in JI,on pharmacokinetics of silibinin. Methods The Beagle dogs received intravenous injection of JI and silibinin,and then its plasma sample was collected in different time. The plasma samples of Beagle were prepared by hydrolysis with sulfatase-? glucuronidase complex enzyme and liquid-liquid extraction with aether. A high-performance liquid chromatographic method was developed to determine the plasma concentration of silibinin,and the pharmacokinetic parameters were performed by BAPP2.3 program. Results The pharmacokinetics of two tested preparations met with two-compartment model. There were not significant differences between pharmacokinetic parameters of JI and that of Silibinin Injection. Conclusion The silibinin in Jiqi Injection has a fast in-vivo clearance rate after intravenous injection,and Panax notoginseng saponins have no effect on its pharmacokinetic parameters.

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

Diagnosis, Differential ; Humans ; Infant ; Male ; Neuroectodermal Tumors, Primitive ; pathology ; Orbital Neoplasms ; pathology

Alzheimer's Disease (AD) is a chronic neurodegenerative disease that usually takes many years from preclinical phase to prodromal phase characterized by mild symptoms before the onset of dementia. Once diagnosed with AD, the brain is already severely damaged and the disease will process quickly to the most severe stages since there is no medications that reverse the neuronal injuries in the brain. Thus, simple, inexpensive, and widely available methods for detecting potential AD patients during their preclinical phases are urgently needed. In such case, olfactory testing may offer a chance for early diagnosis of AD. However, there are limitations in these olfactory tests due to the complexity of the brain areas it extends to and the frequently olfactory fatigue occurred in the behavioral olfactory tests. Great efforts have been done epidemiologically to investigate the correlation between olfactory functions and possibility of developing AD. Different patterns of olfactory dysfunction have been found in AD at early stages and even mild cognitive impairment (MIC), but the cause of the dysfunction remained unclear. Various kinds of AD animal models have been used in the field to clarify the existence of olfactory dysfunctions and thus study the underling mechanism of the dysfunction. In this review we discuss (1) the function of Tau physiologically and pathologically; (2) the genetic background and biological characteristics of the most commonly used Tau transgenic mice; (3) the structural and molecule basis of olfaction; (4) the possible relationship between Tau pathology and olfactory dysfunction. Finally, we suggest that the tau transgenic mouse models may be helpful in studying the possible mechanisms of the dysfunction.
Alzheimer Disease ; physiopathology ; Animals ; Disease Models, Animal ; Mice ; Mice, Transgenic ; Olfaction Disorders ; physiopathology ; tau Proteins

OBJECTIVETo investigate the liver injury in model rats with endotoxemia and to observe the protective effect of Compound 912 Liquid on it.

METHODSRats were randomly divided into three groups, the endotoxemia model group (EMG, injected by lipoplysaccharides (LPS) peritoneally), the intervention group (IG, treated with Compound 912 Liquid via gastrogavage 1 h before model establishing) and the normal control group (NCG). Blood samples of rats were taken at the time points of the 2nd, 4th, 8th, 12th, 48th, 72nd hour and the 7th day after modeling for measuring liver function, levels of plasmatic endotoxin, tumor necrosis factor alpha (TNF-alpha), interleukin-10 (IL-10). The pathological change of liver was observed using light microscope and electro-transmission microscope.

RESULTSThe peak concentration of endotoxin detected at 2 hour after modeling in the IG was significantly lower than that in the EMG (0.358 +/- 0.056 vs 0.685 +/- 0.030), but insignificant difference (P > 0.05) was shown between them in TNF-alpha level. The level of IL-10 continuously rose in IG after treatment, it was still higher than normal level until day 7 (49.096 +/- 4.076 vs 43.454 +/- 5.928, P < 0.05).

CONCLUSIONLPS can induce the increase of serum inflammatory cytokines and anti-inflammatory cytokines in rats to injure liver. Therefore, the inflammatory reaction indicated by LPS may be one of the mechanisms for liver injury. Preventive medication with Compound 912 Liquid showed a significant liver protective effect.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Endotoxemia ; blood ; chemically induced ; drug therapy ; Female ; Interleukin-10 ; blood ; Lipopolysaccharides ; Liver Diseases ; prevention & control ; Male ; Multiple Organ Failure ; prevention & control ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar

An antibiotic-producing bacterium, which was numbered as 20 #-5, was separated from the soil in Chongqing. It was identified as the member of pseudomonas. Gram positive bacteria are badly suppressed by it. The antibiotic secreted by 20 #-5 can endure 100℃ for half an hour, and it can also go through the ultrafiltration membrane with pores of 0.22?m.

Objective To investigate the antiproliferation and induction differentiation of human gas- tric carcinoma which human gastric lower-differentiation mucinous carcinoma MGC-803 cells transplanted in- to nude mice by using rosiglitazone(ROS),and to preliminarily explore the mechanism of differentiation. Methods The mice were randomly divided into five groups:model,ATRA,ROS 25 mg?kg~(-1),ROS 50 mg/kg, ROS 100 mg/kg.After that the volumes were measured and inhibition rates were calculated.The cell cycle was detected by FCM.The protein expression level of Mucin SAC was detected by immunohistochemistry. Results The volume of tumor decreased significantly in ROS treatment groups,the differences had statistical significance compared with model group(P0.05).The xenograft tumors of ROS groups demonstrated the characteristics of differentiation.Xenograft tumor cells were arrested in G_0/G_1 phase,and the cells in S phase decreased significantly,and up-regulated Mucin SAC gene expression.Conclusion ROS could inhibit the growth of tumor,and the effect were dose-dependent with ROS.ROS could induce the differentiation of Xenograft tumor cells of gastric cancer.Its mechanism might be related to the inhibit of transition from G_1 to S phase,degrade the activity of proliferation,regulate the expres- sion of Mucin 5AC.

The applications of targeting gene delivery systems in tumor therapy have attracted extensive attention of researchers in recent years, as they can selectively deliver the therapeutic gene to tumor sites, improve the success rate of gene therapy and reduce the side effects. Therefore, design and development of novel gene delivery vehicles have been a hot area of current research. Recent studies have shown that mesenchymal stem cells (MSCs) have the ability to migrate towards and engraft into the tumor sites. Therefore, these properties make them a great hope for efficient targeted-delivery vehicles in cancer gene therapy. In this review, we examine the promising of utilization of MSCs as a targeted-delivery vehicle for cancer gene therapy, and summarize various challenges and concerns regarding this therapy.
Animals ; Cell Movement ; genetics ; Drug Carriers ; Gene Targeting ; methods ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; physiology ; Neoplasms ; genetics ; therapy ; Recombinant Proteins ; genetics ; metabolism ; Transfection

OBJECTIVETo investigate the nephrotoxic effects of methyl cantharidimide tablets on urinary protein and enzymes in Beagle dogs.

METHODBeagle dogs were randomly divided into negative control group(blank tablet), methyl cantharidimide tablets group (6.11,12.21, 24.42 mg x kg(-1)), continuously 30 days of oral adminiStration, once a day. The drug and control group were collected and determined fresh urine in 1, 2, 3 and 4 weeks of the administration; Serum urea nitrogen (BUN), creatinine (Crea), total protein (TP) and albumin (ALB) as well as sodium, potassium, chloride electrolyte were determined on 15 and 30 days of the administration; Urine albumin (mAlb), kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin( NGAL), N-acetyl-beta-D-glucosaminidase (NAG), clusterin, beta2-microglobulin (beta2-MG), alpha1-microglobulin (alpha1-MG), alanine aminopeptidase( AAP) and im- munoglobulins IgG were tested on 15 and 30 days of the administration.

RESULTCompared with the control group, urine protein and white blood cells was significantly increased in each dose group. On 15 days of the administration, mAlb were higher in each dose group, KIM-1, NGAL, clusterin, NAG and AAP were significantly higher in high-dose group, while the middle and low dose group had no significant difference, as well as blood SCr and BUN no obvious abnormalities. On 30 days, mAlb, KIM-1, clusterin, NAG, AAP were increased in each dose group, appearing dose-effect relationship, beta2-MG and NGAL levels were significantly increased in high-dose group. Contents above indicators were increased with significant dose and time relationship, and serum BUN, Scr were correlated, suggesting that urine mAlb, KIM-1, clusterin, NAG and AAP indicators that can sensitively respond the changes of proteins and enzymes in urine.

CONCLUSIONMethyl cantharidimide tablets has a renal toxicity, urine mAlb, KIM-1, clusterin, NAG and AAP can be used as the early nephrotoxic biomarkers of methyl cantharidimide tablets.

Animals ; Biomarkers ; urine ; Dogs ; Female ; Kidney ; drug effects ; Kidney Diseases ; chemically induced ; Male ; Proteins ; metabolism ; Tablets ; adverse effects ; Urine ; chemistry