BACKGROUND: Embryonic stem cells (ESCs), the seed cells of all mature cells in vivo, are useful tools for nerve transplantation and developmental gene function research. Notch1 signaling pathway is the key pathway to control the ordered neural development and differentiation of many kinds of neural cells, however, there is no report on the dynamic expression of Notch1 signal during the ESC differentiation to date. OBJECTIVE: To investigate the expression of Notch1 protein, transmembrane signal transduction molecule, during directional differentiation of embryonic stem cells into neural cells. DESIGN, TIME AND SETTING: Cell research was carried out between October 2003 and October 2004 at Zhongshan Ophthalmic Center, SUN Yat-sen University, Guangzhou, Guangdong Province, China. MATERIALS: BALB/C mouse embryonic stem cell line Ⅵ (passage 11)was obtained from experimental animal center of SUN Yat-sen University, provided by professor Huang Bing. ESC culture medium was high-glucose DMEM medium with 20% bovine serum and 106 IU/L mouse leukemia inhibitory factor. Induced differentiation medium was high-glucose DMEM medium with 20% fetal bovine.serum and 5×107 mol/L retinoid acid(RA). METHODS: Passage 11 ESCs were resuscitated and incubated by ESC culture medium in incubator at 37℃ with 5% CO2. Passage 11 ESCs were subcultured after 2 or 3 days and RA was added into medium to induce differentiation. Three time points for observation were established: induced for 1, 5 and 9 days. MAIN OUTCOME MEASURES: Morphological changes were observed under inverted phase contrast microscope, MAP-2 antigen expressed in differentiated cells was detected by immunofluorescence method. Immunocytochemistry, Western Blot, flow cytometry assay were used to investigate the Notch1 protein expression. RESULTS: ESCs presented clone-like growth. After induced by RA for 9 days, single neural network was achieved around most of the cell clusters. With the prolongation of induction, MAP-2 positive neural cells increased gradually. Almost all ESC clones expressed Notch1 protein strongly or positively, but Notehl protein expression decreased gradually after induced differentiation (P < 0.01). CONCLUSION: Notch1 signal shuts off progressively during induction of ESCs into neural cells, which suggests Notch1 may play an important role in the differentiation of ESCs into neural cells.

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

Objective We selected baculovirus expression vector system to express human NT-4 with biological activity. Methods The hNT-4 mature peptide-coding sequence is amplified by polymerase chain reaction (PCR), ligated to baculovirus expression vector PacGP67B, and expressed in the insert sf9 cell line. Results The protein molecular weight of the expressed hNT-4 was about 15000 by SDS-PAGE,and the hNT-4 antibody could react with this protein in the infected supernatant and total cell by western-blot . The activity of hNT-4 determined by PC12 cell line was definite. Conclusions The results may aid for studying the effect of the hNT-4 on basic medical research and clinical application.

Objective To improve the understand of the clinical characteristics of mycosis fungoides and provide guidance for the clinical work. Methods The clinical data of 21 patients with mycosis fungoides in our hospital were retrospectively analyzed including the ages at diagnosis, clinicopatholagic characteristics of skin lesions, systemic manifestation, misdiagnosis and treatment of these patients. Results The mean age was (57.3±2.31) years at the diagnosis. Most patients were at the stage of plaques. Clinical manifestations included generalized lesions (52.4 %) and itchy (66.7 %). Epidermotropism (66.7 %) and pautriers microabscesses(57.1%) were common histopathologic features. Previous misdiaguosis rate was 66.7 %. Skin targeted therapies and biologic therapies were effective approaches to relieve the skin rash at early stage, and combined chemotherapy was typically applied in more advanced cases. Conclusion Mycosis fungoides has various clinical characteristics and careful differential diagnosis should be made in clinical practice.

OBJECTIVETo study diffuse alveolar hemorrhage (DAH) in patients with hematologic malignancies after chemotherapy and discuss the possible etiology and appropriate therapy.

METHODSSymptoms, physical examinations, laboratory examination, chest radiographs or computed tomographic (CT) scans, treatments and outcomes of two patients with acute lymphoblastic leukemia (ALL) and non-Hodgkin lymphoma (NHL) each after chemotherapy were presented.

RESULTSBoth of the patients developed cough, progressive dyspnea, a drop of hemoglobin level, hypoxemia and widespread pulmonary infiltrate on chest radiographs or CT scans after chemotherapy. Moreover, case 1 (ALL) had high fever and bloody fluid drained from the intubation of mechanical ventilation, case 2 (NHL) developed continual hemoptysis. They were diagnosed as DAH and improved significantly after intermediate- or high-dose corticosteroid therapy.

CONCLUSIONSDAH is a rare fatal acute noninfectious pulmonary complication in patients with hematologic malignancies after chemotherapy. Early accurate diagnosis, identifying the underlying cause and appropriate treatment are critical for the management of DAH.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Female ; Hemorrhage ; drug therapy ; etiology ; Humans ; Lymphoma, Non-Hodgkin ; drug therapy ; Male ; Methylprednisolone ; therapeutic use ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Pulmonary Alveoli

OBJECTIVETo discuss the clinical effects and superiority of applying external fixation and bone transport to treat osteomyelitis and bone defect of femoral bone.

METHODSFrom August 2008 to December 2013,16 patients with osteomyelitis and bone defect of femoral bone were treated including 11 males and 5 females with an average age of 42 years old ranging from 13 to 62 years old. The average course of disease was 18 months ranging from 2 months to 4.5 years, and the average length of bone defect was 7.8 cm ranging from 4.5 to 15 cm. The bone defect of all cases were treated by external fixation and bone transport, the bone transport began at 1 week after operation, 1 mm per day and 4 times per day.

RESULTSAll patients were followed up for 10 to 36 months (means 22.5 months). One patient did not cooperate with treatment leads to the failure, then took the amputation. The remaining 15 cases of osteomyelitis were under control, including 12 cases of bone transport achieved one stage bone union, 3 cases achieved bone union via bone graft from iliac bone. The bone union time was 5 to 13 months(means 7.9 months). Thirteen patients almost obtained the same length of two lower extremities,2 patients had shortening of 1.5 to 2 cm. The time of moving the external fixation was from 6 to 16 months (means 9.3 months).

CONCLUSIONApplication of external fixation and bone transport is an effective method in treating the osteomyelitis and bone defect that can control the infection, eradicate wounds, and be the equalization of limb length.

Adolescent ; Adult ; Bone Transplantation ; External Fixators ; Female ; Femur ; surgery ; Humans ; Male ; Middle Aged ; Osteomyelitis ; surgery

Angiogenesis Inhibitors ; pharmacology ; therapeutic use ; Animals ; Antimetabolites, Antineoplastic ; therapeutic use ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Deoxycytidine ; analogs & derivatives ; pharmacology ; therapeutic use ; Drug Therapy, Combination ; Female ; Ginsenosides ; pharmacology ; therapeutic use ; Lung Neoplasms ; drug therapy ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation

The potential of Rhizobium sp. N613 to produce the exopolysaccharide (REPS) was studied in this paper. Using an orthogonal design in a flask-shaker culture system, the fermentation medium and conditions of synthesizing REPS were optimized. Based on these results, the fermentation kinetic parameters were obtained in the batch fermentation with a 10L fermentor. The REPS yield of 11.31g/L was achieved by metabolic regulation during 40 h fed-batch fermentation. Transplanted tumor models of sarcoma 180 in mice were used to evaluate the anti-tumor effect. The result of anti-tumor activities showed that inhibition rate was 53.40%, when dose of REPS was 5mg/kg. These results indicate that REPS has the following properties: the short duration of fermentation, the high yield, the low cost, the effective immunocompetence and thickening. Thus, REPS has the value of development and application.

0.05).②There were close relationships between BC047440 gene expression and clinicopathologic findings of liver cancer,including tumor size and portal vein invasion(P

Objective To evaluate the value of stereotactic core needle biopsy(SCNB)in diagnosis of breast lesions.Methods Forty-seven cases were punctured with computer-assisted stereotactic system, spring-loaded biopsy guns and 16 G core needles.The record of each item was collected,including clinical manifestations,descriptions of the mammographic characteristics(such as calcification,mass and architectural distortion),the pathology of the SCNB and the surgical pathology or mammographic follow-up data.Then the results of SCNB were analyzed based on the comparison of SCNB pathology and the surgical pathology.The reason that SCNB failed and misdioagnosed was inferred from the relationship of SCNB accuracy and the X-ray characteristics.Results Forty-four cases were punctured successfully,3 cases failed.Thirty-one patients were operated soon after biopsy.The results of 27 SCNB cases agreed well with the final pathology but the other 4 did not.Conclusions SCNB as an accurate,time-saving and cost- effective method,is also minimally invasive and hardly changes the architecture of the breast.SCNB can diagnose breast lesions in advance,reduces the number of surgical biopsy,and is promising in clinical application.