objective: To offer a model for reference on the operating guideline of ART Institution Ethics Committee to regulate their conducts on ethical supervision in China. Method: Generalizing the mode of committee' action out of duty according to the scientific principle in clause. Conclusion: The suggestion draft, based on committee's practical exploration of ethics, integrates the international practice and external experience for reference properly, which is overall and has stronger operating quality and can draw lessons from the meaning.

Objective To investigate the effects of omeprazole on the expression and the activity of CYP2C19 in HepG2 cells. Methods MTT assay was performed to screen the concentration range of omeprazole which did not inhibit HepG2 cell proliferation. Real-time quantitative polymerase chain reaction(Q-PCR)were carried out to determine the effects of omeprazole (0, 0.1, 1 and 10 mg/L)on the expression of CYP2C19 mRNA, Western blot was carried out to determine the effect of(0, 0.1, 1 and 10 mg/L)on the expression of CYP2C19 protein. After incubation of different concentrations of omeprazole (0, 0.1, 1, 10 and 100 mg/L)with microsomes prepared from insect cells expressing human P450 isozyme, the fluorescence of the samples was detected in a plate reader to analyze if omeprazole inhibited the CYP2C19 activity. Results Omeprazole could down-regulate CYP2C19 mRNA and protein expressions of HepG2 cells in a dose-dependent manner. When the concentration of omeprazole reached 10 or 100 mg/L, the fluorescence intensities obviously decreased, indicating inhibited CYP2C19 activity. Conclusion Omeprazole inhibits the expression and the activity of CYP2C19 in HepG2 cell. When omeprazole is used in combination with drugs activated via CYP2C19 metabolism, their interaction should be considered carefully.

Alzheimer's diseases (AD) is a slowly progressive neurodegenerative disease. The pathogenesis of AD is complex and there is no effective medicine for AD. Catalpol is a kind of iridoid compound, it has neuroprotective, anti-inflammatory and antiaging activities based on cell and animal models of AD. These findings indicate that catalpol may be a potential anti-AD drug. This paper aims to review the advances of catalpol effects on cell and animal models of AD and could provide clue for the research and development of AD drugs.

Objective: To establish an ultra-high-performance liquid chromatograph (UHPLC) method for the determination of paclitaxel (PTX) in polydipeptide paclitaxel (PDP) preparation. Methods: PDP preparation was dissolved in deionized water (DIW) and degraded by 2.0 mol/L sodium hydroxide solution. The concentration of paclitaxel was calculated indirectly by its degradation product. The separation was achieved on an Agilent SB C18 column (2.1 mm × 50 mm, 1.8 μm). Elution was carried out using a mobile phase consisting of acetonitrile-water(10:90, V/V) at the flow rate of 0.2 ml/min. UV detection wavelength was performed at 240 nm and reference wavelength was 360 nm. The temperatures of autosampler and column were thermostated at 15°C (± 0.5) and 40°C (± 0.5°C), respectively. The injection volume was 2 μl. Results: The relationship between the concentration of paclitaxel (0.31-5.00 mg/ml) and the peak area of its degradation product was in good linearity (r = 0.9992, n= 5). Total amount of paclitaxel in different batches of PDP preparation was in the range of 26.77-33.19 mg per vial. Conclusion: The method is accurate, rapid, reproducible and suitable for the analysis of paclitaxel in PDP preparation.

BACKGROUND: Autophagy may be a tumor-inhibiting factor at the early stage of carcinogenesis, while at late stage, it may support or promote the tumor growth and spread, and may contribute to drug resistance. OBJECTIVE: To review the related literature at home and abroad, summarize the relationship between autophagy and osteosarcoma, and explore the mechanism of autophagy in osteosarcoma and its therapeutic prospect. METHODS: CNKI, Wanfang, VIP and PubMed databases were retrieved with the keywords of “autophagy, chemotherapy, autophagy related factors, osteosarcoma, apoptosis, drug resistance” in Chinese and English, respectively. The articles related to the mechanism of autophagy in bone tumors were included. The literature was screened according to the selection criteria, and the repetitive and the long-standing literature were excluded. Fifty-two articles were included for review and analysis. RESULTS AND CONCLUSION: (1) Autophagy may be an important factor in promoting and preventing cancer, and its effect varies with the occurrence of development of tumor. (2) The full understanding and regulation of autophagy pathway can significantly improve the potential of tumor therapy. (3) Considering the importance of autophagy in tumor biology, research on autophagy-regulated miRNA will help us to better understand malignant tumors and may discover new disease markers and treatment strategies. (4) The above conclusion still needs a lot of scientific research and reasonable experiments to confirm.

microRNAs are non-coding microRNAs composed of 18-22 nucleotides, which play an important role in many aspects such as homeostasis regulation and disease development. They have been confirmed to be involved in the pathogenesis of cardiovascular diseases, including atherosclerosis, myocardial infarction, hypertension, and stent stenosis. miRNAs are stable in the blood circulation, which are regulate the proliferation, differentiation, migration, and apoptosis of vascular smooth muscle cells. Apoptosis has special physiological and pathological significance for cardiovascular diseases, so it may be a new hot spot for the diagnosis and treatment of cardiovascular diseases. This article aims to review the association and clinical application of miRNAs in regulating vascular smooth muscle cell apoptosis and cardiovascular disease. It provides a theoretical basis for the research network of miRNAs regulating the function of vascular smooth muscle cells which involved in cardiovascular disease.

Objective: To detect the dynamic change of expression of CD133, a marker of liver cancer stem cell, at different stages of liver carcinogenesis induced by chemicals in C57BL/6J mice. Methods: The tumorigenesis and the growth of chemical (diethylnirtosamine/carbon tetrachloride/ethanol)-induced primary HCC (hepatocellular carcinoma) in 50 male C57BL/6J mice were observed. Another 45 male C57BL/6J mice which had not been exposed to carcinogenic chemicals were used as the normal controls. The mice were randomly sacrificed every two weeks, and the liver tissues were removed to observe the change of pathology. The expression of CD133 mRNA was detected by RFQ-PCR (real-time fluorescent quantitative PCR), and the expression of CD133 protein was detected by immunohistochemistry and Western blotting. The percentage of CD133-positive cells was analyzed by FCM (flow cytometry). Results: The primary HCC was successfully induced in male C57BL/6J mice after chemical intervention for 20 weeks. The results of RFQ-PCR and FCM showed that the expression level of CD133 in the chemicalinduced group was obviously higher than that in the normal control group after 8 weeks (P < 0.05, P < 0.001). The expression level of CD133 kept on rising in the chemical-induced group as time progressed, and it was significantly higher at the 16th week than at earlier stages (P < 0.001). Western blotting result showed that the CD133 weak expression could be observed at the 4th week. As time progressed, the expression level of CD133 protein was gradually increased. The result of immunohistochemistry showed that the expressions of CD133 in the chemical-induced group were weakly, moderately and strongly positive at the 8th, 16th and 20th week, respectively; while the expression of CD133 was negative in the normal control group. Conclusion: The liver cancer stem cell marker CD133 is involved in the development and progression of liver cancer, and its expression level is gradually increased in the process of carcinogenesis. Copyright © 2012 by TUMOR.

OBJECTIVETo investigate the changes and mechanism of angiopoietin1 (Ang1) in murine bone marrow during G-CSF induced mobilization of hematopoietic stem/progenitor cell.

METHODSThe proportion of Lin-Sca1⁺cKit⁺ (LSK) cells in peripheral blood of C57BL/6 mice before and after G-CSF mobilization was detected by flow cytometry. Expression changes of Ang1 and osteocalcin (OCN) during HSC mobilization were determined by immunohistochemistry, enzyme linked immunosorbent assay (ELISA) and real-time fluorescence quantitative PCR. The number of osteoblasts in the bone marrow was counted under the microscope.

RESULTSAfter treated with G-CSF, the proportion of LSK cells in peripheral blood significantly increased from the controls (0.04 ± 0.01)% to (0.61 ± 0.05)% at day 5 (P<0.05). Before G-CSF mobilization, the endosteum cells expressed higher level of OCN and Ang1 than that of bone marrow nucleated cells. The mRNA expression level of OCN was significantly reduced from 28.64 ± 8.61 in the controls to 12.55 ± 7.06 on day 3 and 4.75 ± 1.62 on day 5, and the expression level of Ang1 also declined from 2.84 ± 0.95 in the controls to 0.93 ± 0.30 on day 3 and to 0.92 ± 0.22 on day 5 after G-CSF mobilization. The number of endosteum osteoblasts was significantly decreased after mobilization (P<0.05). The Ang1 expression was decreased in the BM after mobilization. The serum OCN was significantly reduced from (24.11 ± 3.17) ng/ml in the controls to (9.96 ± 2.16) ng/ml on day 3 and (8.43 ± 2.62) ng/ml on day 5, and the Ang1 also declined from (2.24 ± 0.52) ng/ml in the controls to (1.21±0.38) ng/ml on day 3 and (0.90±0.24) ng/ml on day 5.

CONCLUSIONIn G-CSFinduced HSPC mobilization, the bone marrow osteoblasts retraction causes reduction of Ang1, and the reduction of Ang1 may contribute to HSPC mobilization.

Animals ; Bone Marrow ; Bone Marrow Cells ; Granulocyte Colony-Stimulating Factor ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cells ; Mice ; Mice, Inbred C57BL ; Osteoblasts ; RNA, Messenger

Aim To explore the signaling pathway of matrine derivative ZS10 in inhibiting proliferation and inducing apoptosis of BEL-7402 cells. Methods ZS10 was synthesized by organic synthesis. The inhibitory effect of ZS10 on the proliferation of BEL-7402 cells was analyzed by MTT method at the time of 24 h, 48 h and 72 h, respectively, and IC50 was calculated. DAPI staining was used to observe the state of BEL-7402 cells. Clone formation method was used to observe the colony formation of BEL-7402 cells, flow cytometry was used to observe the cell cycle arrest and apoptosis of BEL7402 cells, and Western blot was used to detect the expression level of PI3K/AKT pathway and related proteins. Results MTT results showed that the IC50 was(6.62±1.11)μmol·L-1; DAPI staining showed that the cell state changed significantly with the increase of drug concentration, and the results of colony formation showed that ZS10 significantly inhibited the colony formation of BEL-7402 cells. The results of flow cytometry showed that ZS10 induced S phase arrest and cycle apoptosis of BEL-7402 cells. Western blot showed that ZS10 at the concentration of 08 μ mol·L-1 could regulate the PI3K/AKT pathway and its related proteins in a dose-dependent manner. Compared with the control group, the expression of PI3K, AKT, P-AKT and anti-apoptotic protein Bcl-2 significantly decreased, the expression of pro-apoptotic protein Bax significantly increased, the expression of Cyclin D1 and CDK2 significantly decreased, and the expression of EGFR and N-cadherin, Vimentin significantly decreased in the treatment group. The expression of E-cadherin increased. Conclusions Matrine derivative ZS10 can inhibit the growth and proliferation of hepatocellular carcinoma cell line BEL-7402.

ObjectiveTo design a simple leg-sliding rehabilitation equipment for patients with lower limb dysfunction who need to exercise their leg muscles in bed at the early stage of rehabilitation. MethodsThe mechanism scheme was designed using the crank slider transmission system. The kinematic model was established and the structural dimensions were determined according to the human joint mobility, analysis and verification were then implemented; and the strutural strength of the machanism was verified, and based on the analysis, a protype was built to verify the feasibility of proposed scheme. ResultsThe range of joint activities of the proposed mechanism was in line with the normal human joint activities, the structural strength met the requirements, the prototype operated smoothly, and the actual running speed was basically consistent with the theoretical planning speed. ConclusionThis design could meet the needs of rehabilitation training for bedridden patients with lower limb dysfunction at the early stage of rehabilitation. It has the advantages of small size and light weight, which provides reference value for promoting the development of miniaturized and lightweight lower extremity rehabilitation equipment.