BACKGROUND:Adult central nervous system lacks the ability to regenerate,so it is of great significance to find a new source of neural stem cells.OBJECTIVE:To investigate the differences between basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF)in inducing bone marrow mesenchymal stem cells(MSCs)to differentiate into neurons in vitro.METHODS:MSCs were isolated from normal human bone marrow using density gradient centrifugation and cell attachment method.MSCs were plated in 96-well culture plates at a density of 0.25×108/L and cultured with 200 μL DMEM/F12 for 0,1,2,3,4,5,6,and 7 days,with 20 μL MTT(5 mg/mL)in 5 wells at each time point.The supernatant was removed and 100 μL dimethyl sulphoxide was added to each well for 4 additional hours of incubation.In addition,some cells were exposed to bFGF and EGF.Growth curve was determined with MTT method.Telomerase activity were examined by TRAP(PCR)-ELISA.Additionally,the functional differences of the two cytokines were checked by RT-PCR.RESULTS AND CONCLUSION:RT-PCR revealed that nestin,glial fibrillary acidic protein(GFAP)and neurofilament subunit M (NF-M)mRNA were expressed in un-induced MSCs of passage 4.Nestin expression reduced at 7 days.The expression of micro-tubule-associated protein-2(MAP2)mRNA was not detected until the induction,and increased thereafter.The expression of MAP2 mRNA was greater in bFGF+EGF and bFGF alone groups compared with EGF alone group,and the expression of GFAP in EGF alone group was greater than other groups.Results showed that MSCs can be cultivated,proliferated and differentiated into neural stem cells in vitro.The differentiated neural stem cells have the activity of proliferation,but not have the ability of infinite proliferation as tumor cells.

OBJECTIVE To investigate the changes in etiological factors and the diagnostic methods of fever of unknown origin(FUO).METHODS The clinical data were retrospectively studied from patients with FUO hospitalized in Tongji Hospital from Jan 1992 to Dec 2006.During this time,528 cases fulfilled the criteria of FUO.RESULTS Out of the 528 FUO cases,definite diagnosis was eventually achieved in 476 patients(90.2%).The most common causes of FUO were infectious diseases(54.9%),with tuberculosis accounting for 39.0%(113/290) of cases of infection.Seventy patients(13.3%) were suffered from malignancy;61 patients(11.6%) were suffered from connective tissue disease(CTD);and 55 patients(10.4%) were suffered from other diseases.However,no definite diagnosis had been made in the remaining 52(9.8%) cases until they discharged from the hospital.Pathological diagnosis(30.5%) was the most important diagnostic method,in addition constituent ratio of pathogenic microorganism detection,molecular biology identification technique,immunologic test,diagnostic therapy and imaging accounting for 21.6%,12.8%,12.2%,11.6% and 11.3%,respectively.CONCLUSIONS The causes and diagnostic method of FUO are altered in the past 15 years.Infectious diseases are still the most common cause of FUO,from which tuberculosis is the top one.The cases of fungal infection and AIDS are increased slowly.Malignancy is decreased and occupies the third place of FUO after rheu-matic diseases.Drug induced fever,various kinds of clinical symptoms and syndromes and multifactorial fevers are increased.Diagnostic methods and rate of definite diagnosis are improved.

OBJECTIVE:To discuss the role of hospital pharmacists in the clinical drug trial.METHODS:The role of hospital pharmacists in publicizing and training of Good Clinical Practice(GCP),in the formulation of each standard operating instruction and in each stage of clinical drug trial was analyzed.RESULTS&CONCLUSION:Hospital pharmacists played a key role in clinical drug trial.They can get to know the basic research methodology from participating in the clinical drug trial,meanwhile their consciousness on scientific research can be strengthened.

OBJECTIVE:To develop a RP-HPLC method for simultaneous determination of benzoic acid and salicylic acid in compound benzoic acid ointment.METHODS:Chromatographic condition included Hypersil ODS column and mobile phase of a mixture of potassium dihydrogen phosphate-methanol(78∶22),the detection wavelength was254nm.RESULTS:The linear ranges of benzoic acid and salicylic acid were0.4～2.0mg/ml(r=0.9999)and0.2～1.0mg/ml(r=0.9999);the average re?coveries were98.74%(RSD=0.55%)and98.67%(RSD=0.59%);the intra-day RSDs were0.56%and0.73%(n=5);the inter-day RSDs were0.75%and0.82%(n=5)respectively.CONCLUSION:This method is simple,accurate and quick,and can be used for the quality control of the preparation.

From genomics to proteomics, and then to miRNA, all without exception showed that proteins are the substance which directly regulate life. Comparing with genomics, the study of proteins is more difficult for its great variety, complicated modification and complicated construction. Recently, some new techniques for protein assay and research have provided us, for example, Fluorescence labeling, SELDI, SPR and optical protein chip, of which, SELDI SPR and optical protein chip are label-free. SELDI protein chip technique and its newly developed applications are briefly reviewed here.

OBJECTIVE: To explore and summarize the operation method and operation stage for cochlear implant with chronic suppurative otitis media, to provide the clinical reference for cochlear implant with chronic suppurative otitis media. METHOD: The clinical data of 6 cases of cochlear implant with chronic suppurative otitis media from Jun 2006 to Mar 2009 in our hospital was analyzed retrospectively. The operation stage. surgical skill, possible risk and prognosis was analyzed and summarized. RESULT: 3 of 6 cases received single stage subtotal petrosectomy and cochlear implant. 3 of 6 cases received subtotal petrosectomy, they received staged cochlear implant 1 to 6 months later. No complications occurred, all of the cochlear implanted had good open set speech perception. CONCLUSION Staged operation was the first choice for cochlear implant with chronic suppurative otitis media. Single stage operation took potential risks, it should be done cautiously. The key points for the operation was the clearance of the pathological tissue totally, this required good surgical skill and operation instrument.
Adolescent ; Aged ; Chronic Disease ; Cochlear Implantation ; methods ; Female ; Humans ; Male ; Middle Aged ; Otitis Media ; surgery ; Retrospective Studies ; Treatment Outcome

Immune and hemopoiesis are one of basic project of experimental hematology. Immune function is a essential activity of white blood cells. It was puzzled for the diversity and complexity of immune response. Polarized immune response of immune cells was discovered 30 years ago, which facilitates the study on differentiation of lymphocyte. Recently recognition on multifunctional polarized immune response of lymphocyte and monocyte/macrophage would promote to elucidate the regulatory network of immune cells, diversity and complexity of immune response as well as the study on hemopoiesis. In this paper the approach of multifunctional polarized immune response of lymphocyte, monocyte/macrophage and dendritic cells were reviewed, and their role, especially in cytokine storm and tumor pro-inflammation condition were discussed.
Cell Differentiation ; Cytokines ; immunology ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Humans ; Monocytes ; cytology ; immunology

Objective To study the expression and clinical significance of Notch3 and Notch intracellular domain (NICD) in ovarian carcinoma and the effects of N-[N-(3 ,5-difluorophenyl) acetyl-L-alanyl]-S-phenyl glycine t-butyl ester (DAPT), a γ-secretase inhibitor on the proliferation and apoptosis in OVCAR3, A2780 ovarian carcinoma cell lines. Methods Western blot was used to detect the expression of NICD in the tissues from 58 ovarian carcinomas patients and 21 normal ovarie, who were admitted in Peking University First Hospital from July 2006 to June 2009. Immunohistochemistry was also used to detect the expression of Notch3 in these tissues. The relationship with clinical features of ovarian carcinoma was also analyzed. Proliferation of OVCAR3 and A2780 ovarian cancer cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell cycles and apoptosis and index of proliferation were detected by flow cytometry method. The expression of NICD in OVCAR3 and A2780 cells incubated with DAPT was detected by western blot. Results (1)The expression level of NICD in ovarian carcinomas was significantly higher than that in normal ovarian tissues (1.64 ±0. 19 vs. 0.98 ±0.20;P <0.05). The NICD expression was higher in ovarian cancers with low grade or advanced stage than those in high-middle grade or early stage,respectively (1.90 ± 0. 22 vs. 1.25 ± 0. 21,1.80 ± 0. 21 vs. 1.21 ± 0. 15; all P < 0. 05). The Notch3 protein was stained positively in cytoplasm, nuclear and cell membrane. The expression of Notch3 was higher in ovarian carcinomas than that in normal ovaries [78% (45/58) vs. 24% (5/21); P < 0. 01]. While,there were no stasistical difference in different pathological types, stages, differentiation of ovarian carcinoma. There was no difference between the patients with adjuvant chemotherapy or not. (2)After OVCAR3 and A2780 cells incubated with DAPT 24, 48, 72 hours, NICD expression was significantly lower than that in control group (P < 0. 05). The effects of DAPT inhibited the proliferation and prompted the apoptosis of OVCAR3 and A2780 cells were depended on the concentrations and times. Conclusions Notch3 and NICD may play a key role in the occurrence and progress of ovarian carcinoma. The mechanism of DAPT inhibited the proliferation and prompted the apoptosis of OVCAR3 and A2780 cells may be due to decreased the formation of NICD.

Arrhythmias, Cardiac ; Humans ; Potassium Channels, Voltage-Gated ; genetics

The success of yielding induced pluripotent stem (iPS) cells from human somatic cells demonstrates the important role of reprogramming in the formation of stem/progenitor cells and initiates the exploration of the origin of leukemia stem cells. In our previous work, we have found two types of leukemia, bona fide leukemia and non-bona fide leukemia. Different leukemias originate from different leukemia stem/progenitor cells which are critical to the genesis and evolution of leukemia. Bona fide leukemia and non-bona fide leukemia originate from leukemia stem cells and progenitor cells, respectively. Recent research suggests that different types of leukemia are influenced by the reprogramming state of their origin cells.
Cell Differentiation ; Cellular Reprogramming ; Humans ; Induced Pluripotent Stem Cells ; Leukemia ; genetics ; Neoplastic Stem Cells ; Stem Cells