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 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.

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 study the effect of PCR and bacterial culture in vaginal bacteria test.Methods A total of 84 patients were enrolled in the department of obstetrics and gynecology, affiliated hospital of Nankai university, from February 2014 to March 2016.Vaginal secretion was collected and tested by PCR(group A, n=42), and bacterial culture(group B, n=42).Vaginal bacteria test Resultswere compared in the two groups.ResultsEnterococcus were positive in 13 cases(30.95%)in group A, 3 cases(7.14%)in group B, gardnerella vaginalis were positive in 10 cases(23.81%)in group A, 19 cases(45.24%)in group B;Corynebacterium were positive in 13 cases(30.95%)in group A, 5 cases(11.91%)in group B.Negative rate of vaginal bacterial test was 14.29% in group A, 35.71% in group B.In the above data, the differences between the 2 groups were statistically significant(P<0.05).Conclusion PCR test can effectively improve the positive rate of vaginal bacterial examination, and provide an objective basis for clinical treatment.

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 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.

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.