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.

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 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 observe the clinical effect of low-dose once-daily tadalafil combined with Shuganyiyang Capsules in the treatment of mild-to-moderate erectile dysfunction (ED).

METHODSNinety patients with mild-to-moderate ED were equally randomized to groups A, B and C to receive Shuganyiyang Capsules, tadalafil, and tadalafil + Shuganyiyang Capsules, respectively. The scores of the patients on IIEF-5 and SF-PAIRS (15-Item Short Form of Psychological Interpersonal Relationship Scales) were recorded before and at 1 and 3 months after treatment.

RESULTSThe IIEF-5 scores of groups A, B and C were 10.13 +/- 1.55, 11.00 + 1.60 and 10.73 +/- 1.91 before treatment, and 13.77 +/- 2.11, 17.77 +/- 2.13 and 17.17 +/- 3.84 at 1 month after treatment, significantly higher in B and C than in A (P <0. 001) , but with no remarkable difference between B and C (P =0. 411). At 3 months after treatment, the IIEF-5 scores were 15.77 +/- 2.05, 18.07 +/- 2.24 and 19.37 +/- 3.76 in the three groups, dramatically higher in B and C than in A (P <0.001) as well as in C than in B (P<0.05). The scores on sexual self-confidence, sexual spontaneity and time concerns in SF-PAIRS were 3.90 +/-0.80, 8.67 +/- 1.94 and 14.43 +/- 1.92 before medication, 5.83 +/- 1.02, 9.90 +/- 1.75 and 11.17 +/- 1.68 at 1 month and 6.73 +/- 0.98, 11.07 +/- 2.08 and 10.67 +/-1.60 at 3 months after medication in group A; 4.17 +/- 0.87, 9.37 +/-1.43 and 14.47 +/-1.57 before medication, 6.47 +/-0.78, 10.83 +/- 2.18 and 10.20 +/-1.56 at 1 month and 6.83 +/-0.91, 11.30 +/- 1.88 and 9.47 +/- 1.57 at 3 months in group B; and 4.23 +/-0. 94, 9.50 +/- 1.89 and 14.67 +/- 2.91 before medication, 8.03 +/- 1.67, 13.43 +/-1.10 and 9.70 +/-1.21 at 1 month and 8.93 +/- 1.78, 14.70 +/- 1.26 and 8. 87 +/- 0. 97 at 3 months in group C. Compared with the baseline, the SF-PAIRS scores of the three groups were all significantly improved after treatment (P <0. 05) , and markedly higher in C than in the other two groups (P <0.05).

CONCLUSIONLow-dose once-daily tadalafil combined with Shuganyiyang Capsules is obviously effective in the treatment of mild-to-moderate ED, which not only improves the patients'erectile function, sexual self-confidence and sexual spontaneity, but also reduces their time concerns.

Adult ; Carbolines ; administration & dosage ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Erectile Dysfunction ; drug therapy ; Humans ; Male ; Tadalafil ; Treatment Outcome

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

Analyzed the prescriptions for phlegm retention syndrome that built by Ma Peizhi by the association rules and clustering algorithm, the frequency of drug usage and the relationship between drugs could be get. And from that we could conclude the experiences for phlegm retention syndrome of Ma Peizhi of menghe medical genre. The results of the analysis were that 18 core combinations were dig out, such as Citri Exocarpium Rubrum-Eriobotryae Folium-Citri Reticulatae Pericarpium. And there were 9 new prescriptions were found out such as Aurantii Fructus-Citri Exocarpium Rubium-Eriobotryae Folium-Citri Reticulatae Pericarpium. The results of the analysis were proved that Ma Peizhi of Menghe Medical Genre was good at curing phlegm retention syndrome by using the traditional Chinese medicine of mild and light, such as the medicines of mild tonification, and clearing damp and promoting diuresis.
Algorithms ; Data Mining ; Databases, Factual ; Drug Prescriptions ; standards ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Mucus ; chemistry ; Practice Patterns, Physicians' ; Respiratory Tract Diseases ; drug therapy

The clinical prescriptions of hemoptysis that built by Menghe physician Ma Peizhi were collected, and were analyzed by the method of unsupervised data mining, such as association rules and clustering algorithm. From the prescriptions, we got the frequency of drugs, the association rules among drugs, 10 core drug combinations and 5 new prescriptions. Accordingly, after the analysis, Menghe physician Ma Peizhi had rich experience in the treatment of hemoptysis. His therapy for hemoptysis was clearing heat and moistening lung, dispel heat from blood to stop bleeding, and enriching yin and blood. And we can also make a conclusion that the TCM inheritance support system is of great practical value for minning the old doctors' clinical experiences.
Data Mining ; Databases, Factual ; Drug Prescriptions ; standards ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Hemoptysis ; drug therapy ; Humans ; Practice Patterns, Physicians'

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