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.

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.

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.

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.

Objective To investigate the value of the combined test of five tumor markers for gastric cancer diagnosis.Methods The electrochemical luminescence analyzer was used to measure the serum concentrations of CEA,CA125,CA199,CA724 and AFP in 127 gastric patients and 186 controls,and calculated the sensitivity and specificity.Results The concentrations of CEA (52.9 ±25.5)ng/mL,CA125 (54.2 ±40.6)U /mL,CA199 (42.4 ± 28.8)U /mL,CA724 (9.3 ±6.6)U /mL and AFP (22.6 ±11.4)ng/mL in the gastric cancer group were significantly higher than those in the controls,and the differences were statistically significant (t =6.006,7.118,6.033,6.683, 5.362,all P <0.05 ).The sensitivity in gastric cancer diagnosis with the combined test of five tumor markers (88.2%)was higher than the test of CEA (63.8%),CA125 (59.1%),CA199 (41.7%),CA724 (37.0%)and AFP (46.5%)alone,and the differences were statistically significant (χ2 =20.733,27.754,60.209,71.046, 50.270,all P <0.05).Moreover,the specificity in gastric cancer diagnosis with the combined test of five tumor markers (90.3%)was higher than the test of CA125 (79.3%),while lower than the test of CA724 (97.3%),and the differences were statistically significant (χ2 =9.137,7.832,all P <0.05).Conclusion The combined test of five tumor markers (CEA,CA125,CA199,CA724 and AFP)could increase the rate of gastric cancer diagnosis.

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 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.

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 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.