Based on the analysis on the most common type of scientific research and clinical application of stem cells, including embryonic stem cells , hematopoietic stem cells and mesenchymal stem cells as the breakthrough point, discusses common phenomenon and problems of ethics , scientific research and clinical study the moral life science century new topic, hope scientists, physicians, ethicists, caused by the administrative departments for pub-lic health thought , promote stem cells healthy and steady development .

Objective To determine the protective effect of osteoprotegerin (OPG), receptor activator of nuclear factor kappa-B (RANK) and RANK ligand (RANKL) on avascular necrosis femoral head.Methods Necrotic tissue or corresponding normal tissues were collected from 29 avascular necrosis femoral head patients.Quantitative Real TimePCR ( qPCR ) is used to evaluate mRNA expression of OPG , RANK and RANKL.OPG and RANKL protein levels were estimated by Western blot.Results The results of qPCR showed that the expression of OPG in the necrotic tissue was significantly higher than that in the normal tissue (4.56 ±0.37) (3.39 ±0.52) (P<0.05).The expression levels of RANKL mRNA in necrotic tissues and normal tissues were (0.86 ±0.11) and (0.31 ±0.08), respectively,the difference was statistically significant (P<0.05).The expression levels of RANK mRNA in necrotic tissues and normal tissmes were(0.87 ±0.12), (0.56 ±0.13) respectively,the difference was statistically significant (P<0.05).The ratio of RANKL/OPG and RANK/OPG in normal tissues were 0.69 and 0.52, respectively.In the necrotic tissues, RANKL/OPG and RANK/OPG ratios were 1.35 and 0.61, respectively.Results of Western blot showed that the expression of OPG in necrotic tissues was consistent with that in normal tissues.The expression of RANKL protein was detected in all samples , and the expression of RANKL protein in necrotic tissue and normal tissue was almost the same.RANK protein expression was not detected in all samples.Conclusion OPG, RANK and RANKL play important roles in progress of bone remodeling in necrotic area and in disturbance of bone homeostasis and might have an effect on bone destruction and subsequent collapse of hip joint.

Objective:To explore the therapeutic regimen and key points in the pharmaceutical care for the elderly patients with a-cute exacerbation of the chronic obstructive pulmonary disease ( AECOPD) . Methods: On the basis of clinical pharmacist work, one typical case was selected. Referring to the COPD treatment guidelines, the treatment rationality was analyzed. Meanwhile, an individu-alized pharmaceutical care plan was established and carried out in whole process of the treatment. Results:The pharmacotherapy was effective and rational. By providing the pharmaceutical care for the AECOPD patient, the related problems in the treatment were solved promptly, and the rational advice on the drug treatment was provided. Conclusion:It is very important to enhance integrated pharma-ceutical care in the elderly patients with AECOPD.

Aim To construct a eukaryotic expression vector of human MCHR2 and to analyze the stable expression and signal transduction pathways of MCHR2 in CHO cells. Methods The full-length MCHR2 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and inserted into eukaryotic expres-sion vector pcDNA3.1(+), resulted in the recombinant expression vector pcDNA3.1-MCHR2. The recombinant plasmid was confirmed by the restriction enzymatic digestion and DNA sequencing analysis, and then transfected into CHO cells by lipofectamine. A stably-transfected cell line was obtained by the dominant G418 selection, and the expression of the MCHR2 gene on transcription and translation levels were identified by RT-PCR and Western blot. Signal transduction pathways mediated by MCHR2 were analyzed by measurement of intracellular cAMP and calcium. Results The eukaryotic expression vector pcDNA3.1-MCHR2 was successfully constructed and the MCHR2 gene was stably transfected into CHO cells. A stably-transfected cell line was established and the MCHR2 gene was efficiently transcribed and translated. MCH stimulation had no effect on the production of cAMP, however, it could induce a clear and transient increase of intracellular calcium, suggesting that MCHR2 was only coupled to Gq protein. Conclusion The stable expression of MCHR2 and analysis of its signal transduction pathways in CHO cell line provided a solid experimental foundation for further studies on the function of the MCHR2 gene in vitro.

Objective To investigate the effect and mechanism of oetreotide (OCT) on DENA related hepatoeareinogenesis in rats. Methods Fresh diethylnitrosamine (DENA) solution was given to induce the model of rat hepatoeellular carcinoma. The rats were divided randomly into two groups: OCT treatment group and control group. The survival rate and hepatoeareinogenesis rate were observed. SSTR2 mRNA and protein expression were measured. Results The survival rote of OCT treatment group (70.0%, 7/10) was significantly higher than that of control group (30.0%, 6/20) (X2 = 4.344, P<0.05). 16 weeks after DENA treatment, the difference of bepatoearcinogenesis rate between the two groups was not remarkable though the value of OCT treatment group (0%, 0/10) was lower than that of control groups (30.0%, 6/20)(X2 = 3.750, P＞0.05). However, 22 weeks after DENA treatment, hepatoeareinogenesis in control group (83.3%, 10/12) was markedly higher than that in OCT treatment group (22.2% , 2/9)(X2 =7.843, P<0.01). With liver cirrhosis progressing, the expressions of SSTR2 mRNA and protein increased, and reached the peak 16 weeks after DENA treatment, then began to decrease. The expressions of SSTR2 mRNA and protein in hepatocellular carcinoma were significantly lower than those in the liver 22 weeks after DENA treatment (F = 35.010 and 13. 386, P<0.01). The expression levels in OCT treatment group were similar to those in control group 8 and 16 weeks after DENA treatment. But the expression levels in OCT group 22 weeks after DENA treatment didn't lower markedly, and were higher significantly than those in control group (t = 2.806 and 4.498, P<0.05). Conclusion OCT can inhibit efficiently hepatocareinogenesis and reduce the mortality of rots treated with DENA possibly by a mechanism maintaining the expression levels of SSTR2.

[Objective] Based on the resting state functional magnetic resonance imaging to investigate the abnormal features of the functional connectivity of resting brain neural network in the patients with primary insomnia,by using voxel-wise whole-brain finctional networks analysis of degree centrality (DC) for imaging evidence of neural mechanisms underlying primary insomnia.[Methods] The resting state fMRI were performed in 59 PI patients and 47 age,education,and sex-matched normal healthy subjects.Analysis of DC map changes between the two patient groups and the control group were performed by two sample t test.(threshold at P ＜ 0.05).[Results] Compared with the control group,the patients with PI showed significantly reduced DC value in bilateral medial frontal gyrus (MFG),bilateral anterior cingulate gyrus (ACG),and right insula;and increased DC value in right middle temporal gyrus (MTG),and left cuneus,(CUN),P ＜ 0.05.[Conclusion]Changes of DC value occurred in some region of brain in the P[patient groups when compared with the control group.It was indicated that DC,as a novel resting-state fMRI parameter in the voxel-wise whole-brain functional networks,might be an appealing alternative approach for further study on pathologic and neuropsychological states of PI.

Objective To study the potential effects of angiopoietin 1(Ang1) via adenovirus mediated gene transfer on ischemic heart disease in rabbits. Methods Forty-five male New Zealand white rabbits underwent high positioned double-ligation of the anterior descending left coronary artery, and were divided into three groups: Ang1 group (n=15) received direct myocardium injection of Ang1 recombinant adenovirus; DMEM group (n=15) and LacZ group (n=15) received only DMEM and LacZ recombinant adenovirus as controls respectively. The myocardial infarcted size was evaluated by N-BT macroscopic standing and the degree of angiogenesis was assessed by use of immunohistochemical analysis. The echocardiographic changes were measured on before operation and the 7 th, 14 th, and 28 th postoperative days. Results Animals in three groups had no significant difference in the percentage of infarction size of left ventricle at the 7 th, 14 th day and capillary density at the 7 th day. Animals in Ang1 group showed less infarcted size than DMEM group or LacZ group at the 28 th day and higher capillary density at the 14 th and the 28 th day. The results from echocardiographic measurements showed that animals in all three groups had no significant difference in the left ventricular systolic function before operation and at the 7 th , 14 th day, but the left ventricular systolic function in Ang1 group was better than that in DMEM group or LacZ group at the the 28 th day(P

Epmedii Folium is a commonly used traditional Chinese drug, and is beneficial for the "liver" and "kidney" s function in Chinese medicine. Recently, the origin of this drug is more complex. Most of the identification studies are emphasized on the species certified by the pharmacopoeia and other related species from the same genus of Epimedium, but few was emphasized on the counterfeit. In this paper, one counterfeit of Epmedii Folium, identified as the dried leaf of Quercus variabilis (Fam. Fagaceae), has been reported based on field investigation, comparing specimen of Epmedii Folium and Q. variabilis,using the macroscopic, microscopic and TmC methods. It is resulted that they could be identified clearly not only by the macroscopic features, such as the vein character and the tooth apex, but also by the microscopic features, such as the vascular bundles of the midrib, the non-glandular hair, the anticlinal wall of the epidermis cell and the calcium oxalate crystal. Furthermore their TLC chromatograms showed also difference. This study will give reference for the identification of Epmedii Folium and the related supervision and inspection work.
China ; Discriminant Analysis ; Epimedium ; anatomy & histology ; chemistry ; Plant Leaves ; anatomy & histology ; chemistry ; Plants, Medicinal ; anatomy & histology ; chemistry ; Quercus ; anatomy & histology ; chemistry

This study was performed to prepare immobilized β-glucosidase and snailase, then optimize and compare the process conditions for conversion of icariin. Immobilized β-glucosidase and snailase were prepared using crosslink-embedding method. The best conditions of the preparation process were optimized by single factor analysis and the properties of immobilized β-glucosidase and snailase were investigated. The reaction conditions including temperature, pH, substrate ratio, substrate concentration, reaction time and reusing times of the conversion of icariin using immobilized β-glucosidase or snailase were optimized. Immobilized β-glucosidase and snailase exhibited better heat stabilities and could remain about 60% activity after storage at 4 degrees C for 4 weeks. The optimized conditions for the conversion of icariin were as follows, the temperature of 50 degrees C, pH of 5.0, enzyme and substrate ratio of 1 : 1, substrate concentration of 0.1 mg x mL(-1), reaction time of 6 h for β-glucosidase and 2 h for snailase, respectively. In 5 experiments, the average conversion ratio of immobilized β-glucosidase and snailase was 70.76% and 74.97%. The results suggest an effect of promoted stabilities, prolonged lifetimes in both β-glucosidase and snailase after immobilization. The immobilized β-glucosidase and snailase exhibited a higher conversion rate and reusability compared to the free β-glucosidase and snailase. Moreover, the conversion rate of immobilized snailase was higher than that of immobilized β-glucosidase. The process of icariin conversion using immobilized β-glucosidase and snailase was moderate and feasible, which suggests that immobilized enzymes may hold a promise for industrial usage.
Enzymes, Immobilized ; chemistry ; Flavonoids ; chemistry ; Hydrolysis ; Temperature ; beta-Glucosidase ; chemistry

A study on the microbial transformation of glycyrrhetinic acid (GA) was conducted by a fungus, Cunninghamella blakesleeana CGMCC 3.970 systematically. After incubation with the cell cultures of C. blakesleeana CGMCC 3.970 at 25 degrees C for 7 days on a rotary shaker operating at 135 r x min(-1), GA was converted into one major product and five minor products. The products were extracted and purified by solvent extraction, macroporous adsorbent resin, silica gel column chromatography, and semi-preparative RP-HPLC chromatography. Their structures were identified as 3-oxo-15α-hydroxy-18β-glycyrrhetinic acid(1), 3-oxo-15β-hydroxy-18β-glycyrrhetinic acid (2), 7β,15α-dihydroxy-18β-glycyrrhetinic acid (3), 3-oxo-7β, 15α-dihydroxy-18β-glycyrrhetinic acid (4), 7β-hydroxy-18β-glycyrrhetinic acid(5) and 15α-hydroxy-18β-glycyrrhetinic acid(6) by the analyses of MS, 1H-NMR and 13C-NMR spectroscopic data respectively. Among them, 2 was a new compound. These results suggest that C. blakesleeana CGMCC 3.970 has the capability of selective ketonization and hydroxylation for GA. [Key words] glycyrrhetinic acid; Cunninghamella blakesleeana CGMCC 3. 970; microbial transformation
Biotransformation ; Cunninghamella ; metabolism ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; metabolism ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization