BACKGROUND: Cancer stem cell is a minority population of cancer cells in multiple myeloma possessing the properties of stem cells: self-renewal, multi-directional differentiation and long-term proliferation, which mediating disease initiation, relapse, progression and drug resistance. OBJECTIVE: To review characteristics of biology, phenotypic analyses, sorting and identification in clonogenic myeloma cells, the signaling pathways with in myeloma stem cells and the target therapy related. METHODS: Application of computer search Medline database (1999-01/2009-04), using "multiple myeloma stem cells, heterogeneity, phenotypic analysis, signaling pathways, target therapy" as key words ;application of computer search CNKI database (1999-01/2009-04) and CBM Database (1999-01/2009-04) using "multiple myeloma stem cells, heterogeneity, stem cell separation and identification, signal transduction, targeted therapy as key words" . RESULTS AND CONCLUSION: We collected for 126 literatures on the multiple myeloma stem cell-related, which include 30 Chinese articles and 96 English articles, excluding published earlier, repeated, and similar studies into 30 sub-standard literatures. Now widely recognized that multiple myeloma stem cells may be derived from normal stem cells, the accumulation of mutations and by gene mutation in regaining self-renewal capacity of progenitor cells or fully differentiated mature cells. Circulating clonotypic memory B-cell populations have self-renewal and multi-directional differentiation potential, which represent the cancer stem cells in multiple myeloma. The exact phenotype of multiple myeloma cancer stem cells has not been definitively established and researched remains. At present, both the side population cells and aldehyde dehydrogenase (ALDH) activity assays were mainly capable of isolating multiple myeloma cancer stem cells. Which to possess self-renewal ability by Hedgehog ,Wnt and Notch signaling pathways. When these signal transduction pathways appear abnormal, leading to the occurrence of the tumor and tumor cell growth in non-controlled. Against the cancer stem cell targeted therapy is a new and important direction of selective therapeutic strategies. Cancer stem cell specific surface markers and signal transduction pathways can be used as anti-cancer stem cells to control tumor development targets.

BACKGROUND: Cancer stem cell is a minority population of cancer cells in multiple myeloma possessing the properties of stem cells: self-renewal, multi-directional differentiation and long-term proliferation, which mediating disease initiation, relapse, progression and drug resistance.OBJECTIVE: To review characteristics of biology, phenotypic analyses, sorting and identification in clonogenic myeloma cells, the signaling pathways with in myeloma stem cells and the target therapy related.METHODS: Application of computer search Medline database (1999-01/2009-04), using "multiple myeloma stem cells, heterogeneity, phenotypic analysis, signaling pathways, target therapy" as key words; application of computer search CNKI database (1999-01/2009-04) and CBM Database (1999-01/2009-04) using "multiple myeloma stem cells, heterogeneity, stem cell separation and identification, signal transduction, targeted therapy as key words".RESULTS AND CONCLUSION: We collected for 126 literatures on the multiple myeloma stem cell-related, which include 30 Chinese articles and 96 English articles, excluding published earlier, repeated, and similar studies into 30 sub-standard literatures. Now widely recognized that multiple myeloma stem cells may be derived from normal stem cells, the accumulation of mutations and by gene mutation in regaining self-renewal capacity of progenitor cells or fully differentiated mature cells. Circulating clonotypic memory B-cell populations have self-renewal and multi-directional differentiation potential, which represent the cancer stem cells in multiple myeloma. The exact phenotype of multiple myeloma cancer stem cells has not been definitively established and researched remains. At present, both the side population cells and aldehyde dehydrogenase (ALDH) activity assays were mainly capable of isolating multiple myeloma cancer stem cells. Which to possess self-renewal ability by Hedgehog ,Wnt and Notch signaling pathways. When these signal transduction pathways appear abnormal, leading to the occurrence of the tumor and tumor cell growth in non-controlled. Against the cancer stem cell targeted therapy is a new and important direction of selective therapeutic strategies. Cancer stem cell specific surface markers and signal transduction pathways can be used as anti-cancer stem cells to control tumor development targets.

Objective: To study the effect of PI3K-Akt signaling pathway on the openness degree of mitochondrial permeability transition pore (mPTP)in the aged model rats of myocardial ischemic preconditioning (IPC),and to clarify its possible mechanism.Methods:Thirty-five Wistar Wistar rats (aged 21-23 months)were randomly divided into ischemia reperfusion (I/R)group,I/R+Wort (Wortmannin,inhibitor of PI3K)group,IPC group and IPC+ Wort group (n=7).The models of I/R and IPC were established and 0.6 mg·kg-1 Wort were injected to the caudal veins of the rats in Wort group before reperfusion. After 120 min reperfusion, the myocardium tissue protein was extracted, and the Akt and p-Akt protein expression levels were detected by Western blotting method.The myocardial mitochondrial of rats was separated through differential centrifugation, and the superoxide level, SOD level and openness degrees of mPTP were determined by microplate reader. Results:Compared with I/R group (0.288±0.071),the expression level of p-Akt protein in myocardium tissue of the rats in IPC group (0.346±0.051)was increased (P <0.05),and the expression level of p-Akt im myocardium tissue of the rats in I/R+ Wort group (0.044 ± 0.010)was decreased (P < 0.01).Compared with I/R group (0.216± 0.024),the superoxide level in mitochondrial of the rats in IPC group (0.187 ± 0.022)was decreased (P <0.05),and the superoxide level in mitochondrial in I/R + Wort group (0.218 ± 0.029)had no significant change (P >0.05).Compared with I/R group (1.15±0.15),the SOD activity in mitochondrial of the rats in IPC group (1.39±0.14)was increased (P <0.05),and the SOD activity in mitochondrial of the rats in I/R+ Wort group (1.17±0.21)had no significant change (P >0.05).Compared with I/R group,the mPTP openness degree in IPC group was decreased (P <0.05)during 6 to 2 min,however the openness degree in I/R+Wort group had no significant change (P > 0.05).Compared with IPC group,the mPTP openness degree in IPC+ Wort group was increased (P < 0.05).Conclusion:The IPC of aged rats can inhibit the openness of mitochondrial permeability transition pore through activating the PI3K-Akt signaling pathway.

OBJECTIVE:To establish the technical system that is suitable for protein extracting in Angelicae sinensis seed and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE),and provide technical support for detecting the protein quality and variety purity. METHODS:Using protein content and number of electrophoretic bands as indexes,8 methods,includ-ing voncentrated gel method,salt-soluble protein method,electrode buffer method,dimercaptosyl alcohol (DTT) method,urea method,mercaptoethanol method,trimethylolaminomethane (Tris) method,and acetone precipitation method,were conducted to extract the protein in A. sinensis seed and screen the optimal extraction method. Then based on optimal extraction method,effects of different materials-lipid ratios,sample dilution times(sample volume)and separate gel concentration on SDS-PAGE were investi-gated. RESULTS:Mercaptoethanol method extracted the highest protein contents(29.931 mg/g),with many electrophoretic bands and clear background. When mercaptoethanol method was used as optimal extraction method,electrophoresis effects were the best in the conditions of materials-lipid ratio of 1:10,sample volume of 5 times,separate gel concentration of 15%,which obtained 18 bands totally. CONCLUSIONS:Established protein extraction method and SDS-PAGE technical system are suitable for detecting the purity of A. sinensis seed.

Objective To investigate the effect of tannic acid on glomerular mesangial cells (GMC),and to clarify the mechanism of tannic acid in improving the pathological changes of diabetic nephropathy (DN)from the aspect of oxidative stress and micro-inflammation. Methods The glomerular mesangial cells were treated with glucose (30 mmol·L-1 )or advanced glycosylation end-products (AGEs)bovine serum albumin(BSA)(250 mg·L-1 )and then different concentrations of tannic acid (10,20,40 and 80μmol·L-1 )were added into the GMC.The cells cultured by normal glucose or treated with BSA were used as control groups and then the level of malonic dialdehyde (MDA), glutathione peroxidase (GSH-Px ), superoxide Dismutase (SOD ), CAT (Catalase ) activities and 8-hydroxy-2′-deoxyguanosine(8-OHdG)levels in the culture supernatant 48 h after culture were determined by colorimetry and ELISA method. The expressions of intercellular cell adhesion molecule-1 (ICAM-1 ) protein, monocyte chemotactic protein 1 (MCP-1 ) and ICAM-1 mRNA in GMC were detected by immunohistochemical staining and RT-PCR method.Results Compared with high glucose and AGEs groups,the MDA levels in tannic acid groups were reduced significantly(P<0.05);the activities of GSH-Px,SOD and CAT were increased significantly(P<0.05 or P<0.01);the 8-OHdG levels in annic acid groups were significantly reduced (P<0.05). Compared with high glucose and AGEs groups,the expressions levels of ICAM-1 protein in 40 and 80μmol· L-1 tannic acid groups were decreased (P<0.05 ). The mRNA expressions levels of MCP-1 and ICAM-1 were significantly lower than those in high glucose group (P<0.01 ).Conclusion Tannic acid could protect GMC against the damage of oxidative and inflammatory mediators,thereby delaying and improving the glomerular lesions of DN.

Problem-based learning (PBL) is a student-centered pedagogy in which a subject is approached in the context of realistic problems.We have applied PBL into intemnal medicine of general medicine for up to three years.The results suggest that the PBL method could promote the clinical competencies and self-learning capacities of students.However there is still room for improvement.

Objective To construct a mutant strain of Streptococcus pneumoniae ( S.pneumoniae) with ciaH gene-knockout (ΔciaH) and to analyze the correlation between the ciaH gene and the bacterial re-sistance against β-lactam antibiotics.Methods The ciaH gene segament of S.pneumoniae strain ATCC6306 was amplified by PCR.The PCR product was sequenced after T-A cloning.A suicide plasmid pEVP3ciaH was constructed for the deletion of ciaH gene and then transformed into the ATCC 6306 strain by using the CaCl2 method .The mutant strain of S.pneumonia strain ATCC6306 with ciaH gene-knockout (ΔciaH) was genera-ted through homologous recombination , insertion inactivation and amphemycin screening , which was further identified by PCR , sequencing analysis and laser confocal microscopy .Double agar dilution method was used to detect the minimal inhibitory concentrations ( MICs ) of penicillin G ( PCN ) and cefotaxime ( CTX ) against theΔciaH mutant strain and the wild type strain .The differences between the MICs were further ana-lyzed.The changes of ciaH gene expression at mRNA level after treatment with 1/4 MIC of PCN or CTX were detected by real-time fluorescent quantitative RT-PCR ( qRT-PCR ) .Results The ciaH gene in the genomic DNA of the generated ΔciaH mutant strain was inactivated by insertion as indicated by PCR and se-quencing analysis .Results of the immunofluorescence assay showed that the ΔciaH mutant strain did not ex-press the CiaH protein .The MICs of PCN and CTX against the ΔciaH mutant strain were 32 μg/ml and 64μg/ml, respectively, which were significantly higher than that of the wild type strain (0.06 μg/ml and 1μg/ml) (P<0.01).The expression of ciaH gene at mRNA level was significantly elevated after treatment S.pneumoniae ATCC6306 strain with 1/4 MIC PCN or CTX (P<0.01).Conclusion The CiaH protein in the CiaH/CiaR two-component signaling system is involved in the resistance of S.pneumoniae against β-lac-tam antibiotics.

To investigate the absorption kinetics characteristics of icariin for various intestinal segments.Methods: To establish the situ intestinal perfusion model and to study the absorption of icariin for various intestinal segments.Results: Icariin was metabolized quickly in the four intestinal segments.The permeability coefficient(P*eff) were(4.27?0.28),(5.25?0.17),(1.99?0.09),(0.80?0.03) at duodenum,jejunum,ileum,colon,respectively.Icariside can be detected in the four intestine segments in rats by HPLC.Conclusion: Icariin is metabolized into icariside,and then icariside can be absorbed.

Objective To construct a prokaryotic expression system for expressing the phosphatase-encoding gene phpP in StkP/PhpP signaling couple in Streptococcus pneumonia ( S.pneumoniae) strains, and to further understand the phosphatase activity of the recombinant protein rPhpP.Methods The entire phpP gene of S.pneumoniae strain ATCC6306 was amplified by PCR.The PCR products were sequenced.A prokaryotic ex-pression system for expressing the phpP gene was constructed by the genetic engineering technique.The ex-pressed protein rPhpP and the solubility of rPhpP were assessed by SDS-PAGE and gel image analyzer.Ni-NTA affinity chromatography was performed to purify rPhpP.The changes of phpP gene transcription after the treat-ment with sublethal dosages of penicillin and cefotaxime were determined by real-time fluorescent quantitative RT-PCR.The functional domain in the sequence of the phpP gene and its type was analyzed by bioinformatic softwares.The activity of rPhpP in hydrolyzing the substrate of PP2C phosphotase was measured with Serine/Threonine Phosphotase Assay Kit.The enzyme kinetic parameters of rPhpP were calculated.Results The se-quence of the cloned phpP gene was identical with that reported in GenBank.The rPhpP in soluble form was ex-pressed in the constructed prokaryotic expression system.An increased expression of phpP gene at mRNA level was induced by sublethal dosage of penicillin or cefotaxime.The domain of PP2Cc type phosphatase was detec-ted in the sequence of phpP gene.The purified rPhpP protein could hydrolyze phosphopeptides [ RRA ( pT) VA], a substrate of PP2C type phosphatase, in a dose-dependent manner with Km and Kcat values of 277.35μmol/L and 0.71 S-1 ,respectively.Conclusion The protein encoded by phpP gene of S.pneumoniae was a PP2C type phosphatase.The expression of phpP gene could be enhanced by sublethal dosage of penicillin or ce-fotaxime.

As the new development of scientometrics and informetrics, knowledge graph has infiltrated into the financial, industrial and medical fields, and become a hot issue in the real world research. In this article, the concept and features of knowledge graph, construction and the existing softwares, the application status and development prospect in the TCM field were reviewed, which may provide references for research on the knowledge graph in the TCM field.