Objective To study role of bone marrow mesenehyme stem cells in tumor formation.Methods Nude mice (n = 18) were randomly divided into two groups.Mice in the control group were subcutaneously injected with human embryonic MSCs, and F6 cells were injected into the nine mice of the experimental group.Three mice were sacrificed to remove tumor tissue, which was then prepared for real-time BT-PCR detection at 4 (F6-4), 6 (F6-6) and 7 (F6-7) weeks, respectively.Human lung cancer tissue samples and adjacent non-malignant lung tissue samples were collected from 4 lung cancer patients.The difference between gene expression of F6 cells and MSCs was distinguished by fluorescent differential display (FDD) and verificated by PCR and the western blot analysis. Real-time fluorescent quantitative reverse transcription polymerase chain reaction (FQ-BT-PCR) was used to detect gene expression in tumor tissues after tumorigenesis in nude mice.A new tumor cell line, denominated F6, was established from mutated human embryonic bone marrow mesenchymal stem cells (MSCs) which were induced by the GMCSF and IL-4 in vitro.Results FDD analysis confirmed that cyelin Ⅰ was positively up-regulated in F6 cells compared with MSCs. Similar results were obtained by PCR and western blot.The cyclin Ⅰ gene expression levels in MSCs, F6, F6-4, F6-6 and 176-7 were significantly different(F=12.29 ,P ＜ 0.01).The cyclin Ⅰ gene expression level in F6(4.49±0.40) was 112 folds higher than those of MSCs(0.04±0.02,P＜0.01).Expression levels in F6-4 , F6-6 and F6-7 tissue were 1.82±0.80,3.30±0.43,3.68±1.67 which were significantly higher than that in MSCs (P＜0.05 or ＜0.01).The expression of cyclin Ⅰ increased significantly alone with the accreting volume of tumor.The expression levels of cyelin Ⅰ in human lung cancer tissues in four patients were 0.15±0.02, 0.58±0.23, 4.82±1.12, 1.21±0.60,respectively, and were significantly higher than that in adjacent non-malignant lung tissues (0.04±0.02,0.09±0.04,0.94±0.74,0.15±0.08) (F=12.39,P＜0.01).The protein expression level of cyclin Ⅰ in F6 cells was 0.32±0.08, which was 3.6-fold higher than that in MSCs (0.09±0.06, t=3.86,P＜0.05).Conclusions The expression level of cyclin Ⅰ in tumor tissue is higher during the entire course of tumorigenesis.Cyclin Ⅰ might be one of the factors playing important roles during tumorigenesis,especially in MSCs mutation.

Objective:To investigate whether the IgG from patients with antiphospholipid syndrome(APS) is capable of stimulating blood monocytes to express tissue factor(TF) activity and to explore the mechanisms possibly concerned.Methods:Freshly isolated peripheral blood monocytes were cultured in media containing purified APS IgG or normal IgG or RP-1 [a monoclonal anti-?_2 glucoprotein I(?-?_2GPI)antibody] and/or other agonists and the TF activity on monocytes was investigated by measuring factor Ⅶa-dependent generation of factor Xa.Results:APS IgG as well as RP-1 significantly increased monocytes TF activity,comparing to normal IgG;exogenous ?_2GPI enhanced the effects as the intact IgG molecule.Conclusion:That certain antiphospholipid antibodies,mainly anti-?_2GPI antibodies,induce monocytes TF activity in an antigen-specific manner,not via Fc receptor pathway,is contributed to the thrombotic diathesis in APS.

Objective:To explore the correlation of stress, coping style and the concentration of dehydroepiandrosterone sulfate in umbilical blood between preterm birth and normal birth.Methods:46 with preterm birth and 42 normal birth controls were assessed with Life Event Scale (LES) and Defense Style Questionnaire (DSQ) at 28th-week of pregnancy. The concentrations of DHEA-s in umbilical blood were determined by ELISA for the two groups. Results:Of the scales of malignant life events (frequency: 1.24?0.74 vs 1.04?0.03; strength: 56.21?4.03 vs 44.35?1.06)、immature and middle type defense style scales (4.24?0.13 vs 3.55?0.11; 3.86?0.08 vs 3.64?0.06), and DHEA-s (0.72?0.02 vs 0.33?0.03) there were significant difference between preterm birth and normal birth controls (P

Objective:To investigate the condition of B cell differentiation from cord blood CD34+CD19-hematopoietic stem/progenitor cells in vitro.Methods:CD34+CD19-cells from cord blood were isolated and purified by using immunomagnetic beads separation system.CD34+CD19-cells supported by murine S-17 stromal cells were stimulated in co-culture with T3 and cytokines.Differentiated B cells were analyzed by flow cytometry.Results:The amplification of the B cells derived from CD34+CD19-hematopoietic stem/progenitor cells in co-culture with T3 and IL-7 reached 198-fold of increase,most of the induced cells expressed CD10 and CD19.Conclusion:In the experimental conditions selected,co-culture of CD34+CD19-cells with T3,IL-7 and murine S-17 stromal cells could stimulate differentiate toward to B cells in vitro.

Objective To study the tumorigenesis mechanism in bone marrow mesenchyme stem cells (MSC). Methods The bone marrow MSC could be induced into turnout (F6 cells) in vitro. The difference between gene expression of F6 cells and MSC was distinguished by fluorescent differential display (FDD). Verification of the result was detected by Real time RT-PCR and Western blotting, and immunocytochemistry. Results FDD analysis confirmed that Nucleostemin (NS) was positively up-regulated in F6 cells compared with MSC. Similar results were obtained by PCR and Western blotting. The NS gene expression levels in MSC, F6, 176-4, F6-6 and F6-7 were significantly different(F =160, P <0.05). The NS gene expression level in F6 (0.0372±0.0019) was 18 folds higher than those of MSC(0.0021±0.0002,P <0.05). Expression levels in F6-4, F6-6 and F6-7 tissue were 0.0504±0.0083, 0.0995±0.0026 and 0.0614±0.0036, and were significantly higher than that in MSC(P <0.05). The expression of NS increased significantly with the accreting volume of turnour, and high-level protein expression of NS was confirmed by Western blotting and immunocytochemistry. Conclusion The expression level of NS might be one of the factors playing important roles during turnour genesis, especially in MSC mutation.

BACKGROUND:The microcosmic and submicroscopic organizations of tissue engineering scaffold matedals’superficial structure have all important effect on the eell adhesion and growth.By means of nano.Technique and three-dimensional porous technique,the resultant nano-hydroxyapatite/collagen(n-HAC)call imitate the component and microstructure of natural bone.OBJECTIVE:To observe the biocompatibility of human bone m arrow mesenchymal stem cells(MSCs)cultured in vitro with nHAC.DESIGN,TIME AND SETTING :Single samples observation was performed in the Experimental Center of School ofMedical Technology,Jiangsu University from September 2005 to December 2006. MATERIALD:nHAc was provided by the Material Science and Engineering Department of Tsinghua University.Humanbone marrow mesenchymal stem cells were derived from healthy adult volunteers.All the subiects signed the informedconsents. METHODS:Whole bone marrow culture and successive adherence method was used to culture MSCs in vitro,and the cells were then induced to differentiate into the phenotype of osteoblasts by the revulsants(methylprednisolone,vitamin C,β-glycerophosphate and basic fibroblast growth factor).MSCs at passage 3 were co-cultured with nHACfor 14 days.MAIN OUTCOME MEASURES:The cytological characteristics of the osteoblast were identified throue,alkalinephosphatase immunohistochemistry method and Von Kossa stain.The growth condition with or without nHAC wasevaluated through invert microscope and scanning electron microscope,respectively.RESULTS:The cultured MSCs proliferated into uniform fibroblast-like cells rapidly.MSCs reached confluence and started to form multilayers averaging from 10 to 12 days,passaged stably as well.Then the MSCs passaged from 7 to 9 days.Cytochemistry evaluation showed that MSCs in induced culture were positive for alkaline phosphatase and Von Kossa stain,and deposited calcified matrix.It showed a typical ostcoblast feature in morphology and biology.In coculture model ofMSCs with nHAC,cells would attach to the inner surface of nHAC.At 8 days,the osteoblasts proliferated in the nHAC and the secretion of the matrix was observed.Lots ofcells adheredon the surfaceand pores of nHAC at 14 days.There wereextensive prominent connections among cells. CONCLUSION:THE nHAC is suitable for MSCs to adhere,grow and proliferate,with a good compatibility.

Objective To research the expression and clinical significance of SALL 4 gene in cervical cancer .Methods The ex-pression of SALL4 in 56 samples of cervical cancer and 35 samples of normal cervical tissues was detected by immunohistochemistry and RT-PCR ,and its relationship with the clinicopathological characteristics of cervical cancer was analyzed .Results The expres-sion of SALL4 mRNA was 2 .56 ± 0 .22 in cervical cancer tissues ,which was significantly higher than 0 .38 ± 0 .03 in the normal cer-vical tissues .the difference between them had statistical significance(t=58 .1 ,P<0 .01);the positive expression rate of SALL4 pro-tein was 80 .4% (45/56) in cervical cancer ,which was significantly higher than 11 .4% (4/35) in the normal cervical tissues (χ2 =41 .177 ,P<0 .01) .The positive expression of SALL4 in the cervical cancer tissues was correlated with the differentiation status of tumor ,which in the middle and high differentiation groups was lower than that in the low differentiation group (χ2 =4 .226 ,P=0 .039) ,but had no correlation with age ,International Federation of Gynecology and Obstetrics (FIGO) stage ,tumor size ,pathologi-cal type and lymph node metastasis(P>0 .05) .Conclusion SALL4 is highly expressed in the cervical cancer tissues and correlated with the tumor differentiation ,which might play an important role in the occurrence and development of cervical cancer .

The article is attempted to introduce PBL teaching in clinical biochemistry. The PBL teaching has improved some aspects of students' abilities in the self-study, accessing to information, solving the problem, teamwork and developing thinking. But in the course of PBL teaching, there are three problems: shortage of qualified teachers, teaching system and students' inability. This article puts forward the countermeasures to resolve these problems.

Objective To further define the molecular mechanism involved in the metastasis process in lung cancer and screen out the genes expressed differentially in the lung cancer.Methods mRNA differential display (DD-PCR) was employed to search the specific genes related to metastasis.The highly expressed fragments were cloned and sequenced.Compared with the data in GenBank,the homologous genes were found.The anchor primers were designed to validate the candidates from DD-PCR by real-time PCR.The structure of the gene was prognosticated by software.Results Nine differentially expressed genes were found.One of the nine genes,which named C3orf1,showed high different expression within the six tested cancer cells.The gene was 858bp long and encoded 285 amino acids.The molecular weight was about 32.2 kD.Analyzed by the bio-software,it was found that the gene was consisted with seven EGF-like domains (EGF_1),three 2Fe-2S ferredoxin/iron-sulfur binding regions (2FE2S_FER_1),two VWFC domains (VWFC_1),two thiolase active sites (thiolase_3),and so on.Conclusions The gene C3orf1 is over-expressed in the lung cancer 95D cells.It may encode a familiar secretary growth factor protein and play important role in stimulating growth of the cells.

Objective To screen differentially expressed microRNAs at different stages of hair cycle in a murine model.Methods This study included 30 inbred female C57BL/6 mice (age,6-8 weeks; body weight,15-18 grams).Hair growth cycle was induced in the back skin of C57BL/6 mice by application of wax/rosin followed by depilation under anesthesia witl 1％ chloral hydrate.Three mice were sacrificed by cervical dislocation on day 0,8 and 20 after the induction,and skin tissue was achieved from the same depilated areas parallel to the spine.Total RNAs were extracted from the murine skin and subjected to microarray analysis of microRNA expression.Results Compared with telogen skin,the murine anagen skin showed a higher expression of miR-690,obselote-49 and miR-1308,but a lower expression of miR-291a-5p and miR-212.The expressions of miR-690,obselote-49 and miR-31 were significantly up-regulated,while those of miR-127-3p and miR-212 were downregulated in the catagen skin in comparison with the telogen skin.Conclusion Seven microRNAs are identified in this study to be differentially expressed in murine skin between different stages of hair cycle,which may provide a direction for future research.