Alzheimer's disease (AD) is a chronic neurodegenerative disorder and one of the earliest sings of AD is deficit in short term memory. Till now, the pathogenesis of AD has not been elucidated and the present one-drug-one-target paradigm of anti-AD-drug treatment seems not to be effective in clinic. Multi-target-directed anti-AD-drugs are those agents that may act on two or more targets implicated in AD. Based on the brief introduction of progress in the development of present anti-AD-drugs, the paper mainly focused on the advances in the field of multi-target-directed drug development both home and abroad, especially those studies on selective estrogen receptor modulators.
Alzheimer Disease ; drug therapy ; Animals ; Drug Combinations ; Drug Delivery Systems ; methods ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Indans ; therapeutic use ; Indoles ; therapeutic use ; Selective Estrogen Receptor Modulators ; therapeutic use ; Tacrine ; analogs & derivatives ; therapeutic use ; Thioctic Acid ; analogs & derivatives ; therapeutic use

Transplantation of the microencapsulated recombinant cells is a novel alternative approach to gene therapy of tumors. The semi-permeable membrane of microcapsule protects cells from host's immune rejection, increases the efficiency of gene transfer and reduces the need for frequent injection. Optimization of the preparation and culture is needed to acquire biological microcapsule with high cell viability and protein production. In this work, we studied the effect of different preparation and culture condition on the microencapsulated recombinant CHO cells growth and endostatin production. The result showed that the inoculum cells growth phase and seeding density potently affected the growth and endostatin production of the recombinant CHO cells in the microcapsule. The exponential growth phase recombinant CHO cells with a seeding density of 1 x 10(6) - 2 x 10(6) cells/ mL microcapsules benefited to the cells growth and endostatin production. The time of preparation was another important effect factor of cells viability, the cells viability decreased with the increase of preparation time and the time of preparation should be under 5h for maintaining the cell viability and endostain production. The highest viable cell density and endostatin production was acquired when the microcapsule percentage was 5% in the culture of the microencapsulated cells, the cell growth and endostatin production decreased with the increase of the microcapsule percentage.
Animals ; CHO Cells ; Capsules ; Cell Culture Techniques ; Cell Proliferation ; Cricetinae ; Cricetulus ; Endostatins ; metabolism ; Technology, Pharmaceutical ; instrumentation ; methods ; Time Factors

Community Networks ; Humans ; Public Health

AIMTo study the effects of 1-(2,6-dimethylphenoxy)-2-(3,4-dimethoxyphenylethylamino) propane hydrochloride(DDPH) on brain ischemia injury in rats.

METHODSBy using the middle cerebral artery occlusion (MCAO) induced by nylon surgical thread inserted through the internal carotid artery into the anterior cerebral artery in rats, the effects of DDPH on neuron defects(ND) and infarct size(IS) were investigated. Using incomplete cerebral ischemia in rats, the effects of DDPH on superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in brain tissue and pathological changes in rats were studied.

RESULTSDDPH at the dose of 10 mg.kg-1 i.p. 30 min before ischemia decreased the ND 3 h after ischemia. The IS declined 24 h after ischemia as well. Meanwhile, DDPH was found to increase SOD activity and reduce the MDA content, as well as mitigate pathological damage, of neuron after brain ischemia in rats.

CONCLUSIONDDPH showed protective effects on brain ischemia, probably related to its properties of calcium antagonistic effect and increasing the activity of superoxide dismutases.

Animals ; Brain ; metabolism ; pathology ; Brain Ischemia ; metabolism ; pathology ; Female ; Infarction, Middle Cerebral Artery ; pathology ; Male ; Malondialdehyde ; metabolism ; Neuroprotective Agents ; pharmacology ; Phenethylamines ; pharmacology ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; pathology ; Superoxide Dismutase ; metabolism

The aim of this study was to establish an efficient method for expansion in vitro of natural killer (NK) cells highly purified from human peripheral blood. The CD3-CD56+CD16+ NK cells purified by the negative sorting method of MACS (magnetic microbeads activated cells sorting) were expanded with the different combinations of IL-2, SCF, IL-15 in SCGM (stem cell growth medium) supplemented with 10% human AB serum for 18 days. Cultures were fed with fresh medium and cytokines every 3 days. The sum of cells was counted for evaluating the efficiency of expansion. Then the purity of the CD3-CD56+CD16+ NK cells were determined by flow cytometry and the cytotoxicity to K562 targets was detected by CCK-8 assay in the end. Furthermore, the same way was used to explore the relationship between the efficiency of expansion, cytotoxicity to K562 targets of NK cells and the dose of IL-2. The results showed that after peripheral blood mononuclear cells (PBMNC) were purified by the negative sorting method of MACS, the purity of CD3-CD56+CD16+ NK cells increased from (12.70±2.66)% to (93.03±1.72)%. The CD3-CD56+CD16+ NK cells purified by MACS were expanded with the different combinations of IL-2, SCF, IL-15 in SCGM supplemented with 10% human AB serum for 18 days. The expanding multiple of IL-2/IL-15/SCF group was significantly higher than other groups (p<0.05). The purity of NK cells in the groups with cytokines was not significantly lower than that before expansion (p>0.05). The cytotoxicity of the groups with cytokines was significantly higher than that before expansion. Especially, the cytotoxicity (%) of NK cells in IL-2/IL-15 group and IL-2/IL-15/SCF group was more than 90%. The expanding multiples of low-dose group, medium-dose group and high-dose group were significantly higher than that of zero-dose group (p<0.05), but no significant difference was found between themselves (p>0.05). The cytotoxicity of the groups with IL-2 was significantly higher than that before expansion. Cytotoxicity to K562 cells in high-dose group was significantly higher than that in others (p<0.05); there was no significant difference between low-dose group and medium-dose group (p>0.05). It is concluded that cytokines in the 4 groups were efficient for expansion and the cytotoxicity of highly purified NK cells in vitro. IL-2/SCF/IL-15 combination is the most efficient one among different combinations, and enhanced significantly the cytotoxicity of NK cells against K562 targets. The efficiency of expansion and the cytotoxicity in vitro of NK cells are not related with the dose of IL-2, when IL-2<1,000 U/ml. It is indicated that IL-2 of high-dose (≥1,000 U/ml) may enhance the cytotoxicity of NK cells in vitro more efficiently.
CD3 Complex ; immunology ; CD56 Antigen ; immunology ; Cell Culture Techniques ; methods ; Cell Separation ; methods ; Cells, Cultured ; Humans ; Immunophenotyping ; Interleukin-2 ; pharmacology ; K562 Cells ; Killer Cells, Natural ; cytology ; immunology ; Receptors, IgG ; immunology

Adult ; Aged ; Bone Transplantation ; adverse effects ; Female ; Follow-Up Studies ; Humans ; Ilium ; surgery ; Male ; Middle Aged ; Postoperative Complications ; pathology ; prevention & control ; therapy

This study was aimed to investigate the effects of focal adhesion kinase (FAK) gene silence on leukemia cell growth, leukemogenesis and efficacy of chemotherapy drug. Vector containing lentiviral-FAK-shRNA was constructed and transfected into BCR/ABL-BaF3 leukemic cells, the cell growth and apoptosis were detected in vitro. The effect of FAK shRNA on leukemogenesis was studied in a murine model with leukemia. The apoptosis of leukemia cells and survival of leukemic mice treated by FAK shRNA combined with drug STI571 were monitored. The results showed that FAK gene expression was knocked down by lentiviral-FAK-shRNA. FAK gene silencing inhibited leukemia cell growth in vitro. The apoptosis test results showed that the percentages of Annexin V(+) cells in vector control group and FAK shRNA group were (3.46 ± 0.56)% and (7.3 ± 0.79)%, respectively, and the difference was statistically significant (p < 0.05). The mice in vector control group died at day 21 to 27, while the mice in FAK shRNA group died between day 52 and 60, and the difference was statistically significant (p < 0.05). Moreover, FAK gene silence combined with drug STI571 could enhance the apoptosis of leukemia cells and prolong survival time of leukemic mice. It is concluded that FAK gene silence inhibits leukemogenesis and promotes efficacy of chemotherapy drug on leukemia cells, indicating FAK gene silence may be considered as a new therapeutic strategy for leukemia.
Animals ; Focal Adhesion Kinase 1 ; genetics ; Gene Silencing ; Genetic Vectors ; Leukemia, Experimental ; genetics ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo study the correlation between brain edema, elevated intracranial pressure (ICP) and cell apoptosis in traumatic brain injury (TBI).

METHODSIn this study, totally 42 rabbits in 7 groups were studied. Six of the animals were identified as a control group, and the remaining 36 animals were equally divided into 6 TBI groups. TBI models were produced by the modified method of Feeney. After the impact, ICP of each subject was recorded continuously by an ICP monitor until the animal was sacrificed at scheduled time. The apoptotic brain cells were detected by an terminal deoxynucleotide-transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay. Cerebral water content (CWC) was measured with a drying method and calculated according to the Elliott formula. Then, an analysis was conducted to determine the correlation between the count of apoptotic cells and the clinical pathological changes of the brain.

RESULTSApoptotic cell count began to increase 2 h after the impact, and reached its maximum about 3 days after the impact. The peak value of CWC and ICP appeared 1 day and 3 days after the impact, respectively. Apoptotic cell count had a positive correlation with CWC and ICP.

CONCLUSIONSIn TBI, occurrence of brain edema and ICP increase might lead to apoptosis of brain cells. Any therapy which can relieve brain edema and/or decrease ICP would be able to reduce neuron apoptosis, thereby to attenuate the secondary brain damage.

Animals ; Apoptosis ; Brain Edema ; etiology ; metabolism ; pathology ; Brain Injuries ; complications ; pathology ; physiopathology ; Cell Count ; Disease Models, Animal ; In Situ Nick-End Labeling ; Intracranial Hypertension ; etiology ; pathology ; physiopathology ; Male ; Necrosis ; genetics ; pathology ; Rabbits ; Reference Values ; Telencephalon ; metabolism ; Water ; metabolism

BACKGROUNDInsulin-like growth factor-II mRNA-binding protein 3 (IMP3) is a newly identified mRNA-binding protein that is involved in embryogenesis and carcinogenesis of some malignant tumors. The aim of this study was to detect the expression of IMP3 protein in gastric adenocarcinoma (GAC) and the correlation with clinicopathological features.

METHODSIMP3 protein in 92 samples of GAC was evaluated by immunohistochemical method. The Mann-Whitney U test and Kruskal-Wallis H test were used to compare IMP3 expression and clinicopathological parameters. Kaplan-Meier survival curve, log-rank test and Cox-regression model were used to evaluate the correlation of IMP3 protein expression to the prognosis of patients.

RESULTSOut of 92 cases of adjacent normal mucosa (ANM), 10 with dysplasia demonstrated weak expression of IMP3 and 82 without dysplasia showed negative expression. Out of 92 cases of GAC, positive immunohistochemical stain for IMP3 was identified in 75 (82%) cases. A comparison of IMP3 expression in GAC and ANM showed stronger immunohistochemical reactivity in GAC (P < 0.05). High expression of IMP3 was found to be associated with lymphoid metastasis, high Ki-67 labelling index, and patient poor outcome (P < 0.05). There was a significant TNM stage difference between GAC with and without IMP3 expression (P < 0.05). Tumors with higher stage showed higher level of IMP3 expression. In multivariate analysis, IMP3 emerged as an independent predictor of survival.

CONCLUSIONSIncrease of IMP3 expression suggests that IMP3 may play an importent role in the carcinogenesis and tumor metastasis in GAC. It could be regarded as a novel proliferation and prognostic indicator for patients with GAC.

Adenocarcinoma ; chemistry ; mortality ; pathology ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; analysis ; Female ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; analysis ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Prognosis ; RNA-Binding Proteins ; analysis ; physiology ; Retrospective Studies ; Stomach Neoplasms ; chemistry ; mortality ; pathology

OBJECTIVETo improve Madigan prostatectomy (MPC) for a much satisfactory effect in open surgery.

METHODSA total of 52 patients with benign prostatic hyperplasia (BPH) were treated using MPC. The MPC procedure was modified by exposing anterior prostatic urethra near the bladder neck and conjunction with cystotomy. This modified procedure preserved prostatic urethra intact and could also deal with intracystic lesions at the same time.

RESULTSThe intact of prostatic urethra was kept completely or almost for 48 cases. The hemorrhage amount during modified procedure was a less. The mean operative time was 120 minutes. The 35 patients had been followed up for 1 - 12 months. The average Qmax was 18.9 ml/s. The cystourethrography revealed that the urethra and bladder neck were intact in 8 patients postoperatively. Furthermore, the prostatic urethra was obviously wider after modified MPC.

CONCLUSIONSThe modified MPC can reduce the urethra injury and enlarge the MPC indications. The modified technique is easy to perform with little complications and much more satisfactory clinical result. The modified MPC is highly recommended.

Aged ; Humans ; Male ; Middle Aged ; Prostatectomy ; methods ; Prostatic Hyperplasia ; surgery