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 correlation between amino terminal B-type natriuretic peptide precursor(NT-proBNP) and risk factors of acute coronary syndrome (ACS) with elder patients.Methods One hundred and twenty-eight ACS patients were divided into unstable angina pectoris (UAP) group with 52 samples,ST elevation myocardial infarction (STEMI) group with 35 samples and non-ST elevation myocardial infarction (NSTEMI) group with 41 samples.Meanwhile 45 healthy elder people were adopted as control group.Firstly,the subjects of blood pressure,body mass index (BMI) and smoking numbers were measured.Secondly,venous blood was collected to assay NT-proBNP,cardiac troponin Ⅰ (cTn Ⅰ),homocysteine (Hcy),blood-lipoids and C-reactive protein(CRP).Lastly,ultrasonic cardiogram was used to test left ventricular end diastolic diameter(LVEDD),left ventricular end systolic diameter (LVESD) and left ventricular ejection fraction (LVEF).In addition,correlation analysis was researched between NT-proBNP and other factors.Results NT-proBNP levels of UAP,STEMI and NSTEMI groups were significantly higher than the control group ((794.18±182.64) ng/L,(872.43±245.67) ng/L,(557.25±163.81) ng/L) and (125.84±59.27) ng/L,P ＜ 0.05).NT-proBNP was positive correlation with systolic blood pressure,diastolic blood pressure,Hcy and CRP (r=0.182,0.176,0.281,0.191;P=0.040,0.043,0.001,0.031),however negative with LVEF(r=-0.247,P =0.005).Conclusion NT-proBNP level is sensitive to monitor ACS variety,and it is significant to test NT-proBNP combining Hcy,CRP,and cTn Ⅰ for diagnosing and treating ACS.

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.

According to the three orientation cultivating mode,in order to improve quality of experiment teaching in pathogen biology and cultivate innovative talents of medicine the open experi-ment teaching which based on the research projects is applied to optimize the experiment content design, teaching form and comprehensive assessment. For example, In the choice of subject direction more attention are paid to the students' autonomy; In the design of experiment content students' inter-ests should be fully considered;During the course of experiments, the teachers play the role of visitors most of the time; In the experiment evaluation, students' ability of solving problems and their innova-tive ability are included. By participating in research projects, it not only can arouse the enthusiasm of students, but also can cultivate their scientific spirit of innovation and the rigorous scientific attitude.

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.

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.

ObjectiveTo establish a analytical system for the survival motor neuron (SMN) subtle mutation,and evaluate its application in two families with spinal muscular atrophy (SMA).MethodsSMN genes in seven family members from two SMA families were analyzed at both transcript level and genomic level,by the use of the conventional PCR-RFLP,allele-specific PCR,multiplex ligation-dependent probe amplification (MLPA) and T subcloning and sequencing of SMNI gene.ResultsIn family A,the patient had a single SMN1 copy who was carrying nonsense mutation L228X,which was also found in his father.In family B,as the patient's sample was unavailable,the father was indeed a carrier with one normal SMN1 allele and the other SMN1 allele carrying a frameshift mutation 22_23insA.The remaining family members were SMA carriers with one SMN1 copy.ConclusionThis analytical system for SMN subtle mutation offers viable molecular basis for genetic counseling and prenatal diagnosis in SMA families.

BACKGROUND:Previous research has shown that stem cells at different age stages had various proliferation and differentiation in vitro.Therefore,biological characteristics of bone marrow-derived mesenchymal stem cells at different age stages play a key role in evaluating differentiation into myocardial cells and determining proliferation and differentiation parameters in vitro.OBJECTIVE:To compare in vitro proliferation,surface marking,and differentiation into myocardial cells of bone marrow-derived mesenchymal stem cells from different aged rats.DESIGN,TIME AND SETTING:An in vitro contrast cytological observation was performed at the Laboratory of Biochemistry and Molecular Biology,Shanxi Medical University from May 2008 to May 2009.MATERIALS:A total of 15 healthy male Wistar rats were divided into three groups:1-month-old,3-month-old,and 12-month-old groups,with 5 rats in each group.METHODS:Bone marrow-derived mesenchymal stem cells were cultured using adherence screening method.At the third passage,cells differentiated into myocardium-like cells by the induction of 5-azacytidine.MAIN OUTCOME MEASURES:Cell proliferation was observed using MTT;cell surface marking was detected using flow cytometry;cardiac-specific myosin heavy chain mRNA expression was measured using RT-PCR.RESULTS:Bone marrow-derived mesenchymal stem cells were adherent in each group,and the proliferation curve was similar to each other.Under the same culture condition,the proliferation in the 1-month-old and 3-month-old groups was faster than 12-month-old group (F=28.71,P ＜ 0.05).The third-passage cells strongly expressed CD44 and CD71 but weakly expressed CD34 and CD45.At 2 weeks after 5-azacytidine induction,cardiac-specific myosin heavy chain mRNA was observed in all groups;however,the cardiac-specific myosin heavy chain mRNA expression in both 1-month-old and 3-month-old groups was significantly greater than 12-month-old group (t=5.140,2.827,P ＜ 0.05).CONCLUSION:With the age increasing of rats,proliferation,survival time,and differentiation of bone marrow-derived mesenchymal stem cells were weakened.

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.