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.

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.

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 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.

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 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.

Objective To analyze the clinical data of recipients over 15 years after renal transplantation,and to find the factors that affect the long-term survival of recipients after renal transplantation.Method Before June 30,2000,326 renal transplant recipients in our hospital were collected retrospectively.The risk factors which affect the survival of kidney transplant recipients and kidney were analyzed from four dimensions.A Cox model was established to analyze these multi factors.Result Cox hazard model indicated that advanced age (P=0.010,RR =1.052),AMR (P＜0.001,RR =18.311),nonadherence (P =0.001,RR =2.854),smoking (P =0.025,RR =2.097)were the risk factors for recipients' survival.Using immunosuppressive regimen FK506 + MMF+ Pred (P =0.019,RR =0.433),or CsA + MMF + Pred (P =0.019,RR =0.413) was the protective factor for recipients' survival.Nonadherence (P＜0.001,RR =5.645),and diabetes (P＜0.001,RR =3.310) were the risk factors of grafts' survival.Using immunosuppressive regimen FK506 + MMF + Pred (P＜0.001,RR =0.236),or CsA + MMF + Pred (P =0.002,RR =0.317) was the protective factor of grafts' survival.Conclusion To enhance the long-term outcome of recipients and grafts,the individualization of immunosuppressive regiments and controlling of the chronic diseases progress by changing the unhealthy life style are cutting on edge.

Objective To investigate the relationship between bone mineral density (BMD) and the changes of serum adiponectin (APN) level in elderly men.Methods A total of 240 elderly men was enrolled in this study.Measurement of BMD of lumbar spine with dual energy X-ray absorptiometry was performed.All subjects were divided into three groups (normal,osteopenia,and osteoporosis) according to the T value of BMD.Serum APN level was tested using enzyme-linked immunosorbent assay (ELISA),meanwhile were recorded as bone lurnover markers.Results (1) The level of serum APN in osteopenia group was lower than in normal group,the level of serum APN in osteoporosis group was significantly lower than in osteopenia group,while the level of serum APN in osteoporosis group was significantly lower than in normal group (all P ＜ 0.05).(2) Serum APN was positively correlated with T value of BMD (r =0.475,P ＜0.01).(3) The stepwise multiple linear regression analysis showed serum APN,25-hydroxyvitamin D,β-isomerized carboxyterminal propeptide (β-CTX),and osteocalcin could enter into the equation.Conclusions Serum APN might play an important role in pathogenesis of osteoporosis in elderly men.