BACKGROUND: Glial cell line derived neurotrophic factor (GDNF) plays an important role in inducing the differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro and promoting the regeneration of neuron axons. OBJECTIVE: To observe BMSCs differentiation induced by over-expression of GDNF gene, detect synaptic function of cells and expression of Wnt signaling pathway components after differentiation, and preliminarily explore the mechanism of BMSCs differentiation into mature neurons. METHODS: Rat BMSCs were isolated and cultured, and further divided into recombinant adenovirus-containing GDNF gene transfection group (Ad-GDNF-BMSCs), adenovirus transfection control group (Ad-BMSCs), and untransfected control group. The relative expression of GDNF gene in BMSCs of each group was detected by Q-PCR, and the expression of β-catenin, cyclin D1, NeuN and MAP-2 was detected by immunofluorescence technology in each group. High K+ stimulation induced cell depolarization response after differentiation, and FM4-64 marks synaptic vesicle activity of differentiated cells. RESULTS AND CONCLUSION: (1) The adenovirus-transfected gene had no significant negative effect on the proliferation of BMSCs. BMSCs could express endogenous GDNF gene continuously and at high levels. (2) Under the induction of GDNF gene, BMSCs could express neuron-specific protein NeuN after 3 days cultivation in vitro. The expression of β-catenin protein also could be detected in the cytoplasm of cells. After 7 days cultivation in vitro culture, BMSCs expressed the mature neuronal marker protein MAP-2, and the cell body contracted significantly. Neuron axon-like structures appeared around the cell body. Moreover, β-catenin and cyclin D1 were respectively detected in the cell cytoplasm and the nucleus, while the expression levels of NeuN, MAP-2, β-catenin, and cyclin D1 were not observed in Ad-BMSCs and untransfected control groups, and the cells remained spindle-shaped. (3) After 11 days of in vitro culture, the cells in the Ad-GDNF-BMSCs group showed typical neuronal processes or axons and connected to each other into a network structure, which could be labeled with FM4-64 to show red fluorescence, and induced by high K+ stimulation to induce action potentials in the cells. Synaptic vesicle activity in posterior axons showed FM4-64 red fluorescence gradually decaying. Under the same conditions, cells in the Ad-GDNF-BMSCs group and untransfected control group did not present FM4-64 fluorescently labeled synaptic vesicle activity. (4) Continuous GDNF induction can promote BMSCs differentiated into mature neurons with synaptic cycle function, and may be carried out through the classic Wnt/β-catenin signaling pathway.

To study the in vitro anti-angiogenesis effect of three curcumin pigments (curcumin, demethoxycurcumin, bisdemethoxycurcumin). In the study, the inhibitory effect of the three curcumin pigments on proliferation of HUVEC cells induced by OX-LDL and the effect on migration of HUVEC cells were detected. The effect on neovascularization was observed by chorioallantoic membrane (CAM) test. The effect on cell adhesion factors ICAM-1 and VCAM-1 of HUVECs were tested by Real-time RT-PCR. It was found that the three curcumins could inhibit the proliferation of HUVEC cells induced by OX-LDL within the dosage range 4, 8, 16 mg x L(-1), with a dose-dependence. The proliferative effect of curcumins on HUVECs was greater than the other two derivatives (P < 0.01). All of the three curcumin pigments inhibited the migration of HUVEC cells and the angiogenesis of chick chorioallantoic membrane (CAM). The migration inhibition rate of curcumins at middle and high concentrations was greater than the other two (P < 0.01). All of the three curcumin could down-regulate the expression of VEGF and ICAM-1, and curcumins showed more obvious effect in down-regulating VEGF than demethoxycurcumin and bisdemethoxycurcumin(P < 0.01); Bisdemethoxycurcumin showed the most significant effect in down-regulating ICAM-1 (P < 0.01). All of the three showed no remarkable effect on expression of VCAM-1, and only bisdemethoxycurcumin showed the down-regulating effect (P < 0.05). According to the findings, all of the three curcumin pigments could resist angiogenesis by inhibiting proliferation and migration of endothelial cells and down-regulating the expression of VEGF and adhesion molecules ICAM-1.
Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Movement ; drug effects ; Cells, Cultured ; Chorioallantoic Membrane ; drug effects ; Curcumin ; analogs & derivatives ; pharmacology ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; RNA, Messenger ; analysis ; Vascular Cell Adhesion Molecule-1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics

A novel method was developed for the rapid determination of multi-indicators in corni fructus by means of near infrared (NIR) spectroscopy. Particle swarm optimization (PSO) based least squares support vector machine was investigated to increase the levels of quality control. The calibration models of moisture, extractum, morroniside and loganin were established using the PSO-LS-SVM algorithm. The performance of PSO-LS-SVM models was compared with partial least squares regression (PLSR) and back propagation artificial neural network (BP-ANN). The calibration and validation results of PSO-LS-SVM were superior to both PLS and BP-ANN. For PSO-LS-SVM models, the correlation coefficients (r) of calibrations were all above 0.942. The optimal prediction results were also achieved by PSO-LS-SVM models with the RMSEP (root mean square error of prediction) and RSEP (relative standard errors of prediction) less than 1.176 and 15.5% respectively. The results suggest that PSO-LS-SVM algorithm has a good model performance and high prediction accuracy. NIR has a potential value for rapid determination of multi-indicators in Corni Fructus.
Algorithms ; Calibration ; Cornus ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Least-Squares Analysis ; Models, Theoretical ; Neural Networks (Computer) ; Quality Control ; Spectroscopy, Near-Infrared ; Support Vector Machine

OBJECTIVETo investigate the role of non-high density lipoprotein cholesterol (non-HDL-C) in the assessment of cardiovascular disease (CVD) risk factors such as hypertension, pre-diabetes and diabetes in obese children.

METHODSAccording to the presence of complications (hypertension, pre-diabetes and diabetes), 810 children with central obesity were divided into two groups: one group with complications (n=499) and one group without complications (n=311). One hundred and sixty-four age- and sex-matched children served as the control group. Logistic regression analysis and receiver operating characteristic (ROC) curves were used to analyze the detection of non-lipid CVD risk factors by seven lipid markers.

RESULTSThe prevalence rates of hypertension and pre-diabetes were significantly higher in obese children with high non-HDL-C concentrations (≥3.76 mmol/L). After adjusting for waist circumference Z-scores, the area under the ROC curve for non-HDL-C was 0.680 to detect non-lipid CVD risk factors, while the areas for low-density lipoprotein cholesterol, total cholesterol and apoprotein B were 0.659, 0.669 and 0.647 respectively.

CONCLUSIONSCompared with the other lipid markers, non-HDL-C is a better predictor for non-lipid CVD risk factors in obese children. Measurement of non-HDL-C concentations is recommended for obese children.

Adolescent ; Cardiovascular Diseases ; etiology ; Child ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Female ; Humans ; Logistic Models ; Male ; Obesity ; blood ; complications ; Risk Factors

The study was aimed to investigate the immunophenotypic and cytogenetic features of chronic lymphocytic leukemia (CLL) in order to provide an evidence for diagnosis and therapy. Immunophenotypic analysis was performed by using a panel of monoclonal antibodies and three-color immunofluorescence staining methods of flow cytometry in 51 patients with CLL, and the cytogenetic features were analyzed by R-banding technique. The results indicated that among 51 CLL cases, the positive rate of CD19 and CD23 was 96.1%, followed by CD15 (94.1%), CD20 (82.4%) and CD22 (78.4%). The positive rate of CD38 was 23.5%. Forty-six patients expressed both CD5 and CD19. Seven main clonal chromosomal abnormalities were detected by conventional cytogenetics (CC) in eighteen cases (35.3%), with three cases of +12, two cases of 13q(-), other chromosomal abnormalities included +14, 6q(-), t (11; 14), t (14; 18) and t (2; 7). Expression of the antigens had no relationship with chromosomal abnormalities. It is concluded that typical CLL express CD5, CD19 and CD23, and the positive rate detected by CC in CLL is low. Immunophenotyping in combination with cytogenetic technique plays an important role in the diagnosis and prognosis of CLL.
Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; Antigens, CD ; metabolism ; Antigens, CD19 ; metabolism ; Antigens, CD20 ; metabolism ; Chromosome Aberrations ; Chromosomes, Human, Pair 12 ; Chromosomes, Human, Pair 13 ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; immunology ; Lewis X Antigen ; metabolism ; Male ; Middle Aged ; Translocation, Genetic ; genetics

OBJECTIVETo investigate the effects of top pruning on fruiting characters of Platycodon grandiflorum, and find the suitable stage, in which seed growth and development furtherly.

METHODOne-year old seedlings were chosen and planted in field. Plant height, branching number, fruit number per plant, 1000 grains weight were measured during growth and development period, respectively.

RESULTThe treatment of top pruning postponed in turn the flowering date, lowered the plant heights and the fruit number per plant, increased the branching number and influenced significantly on 1000 grains weights.

CONCLUSIONThe suitable stage of top pruning for producing seeds was from June 20th to July 5th.

Crops, Agricultural ; growth & development ; Fruit ; growth & development ; Platycodon ; growth & development

OBJECTIVETo assess the therapeutic effect and safety of electroacupuncture (EA) at Zhigou (TE 6) on constipation induced by stagnation or deficiency of qi.

METHODSMulti-central randomized controlled trials were adopted and the patients conforming to the criteria for diagnosis, enrolling and excluding were divided into an observation group and a control group according to random number table. The observation group were treated with EA at Zhigou (TE 6) and the control group with EA at non-acupoint for 4 weeks. The cumulative score of clinical symptoms, colon transmission test and various safety indexes were assessed. The following-up was carried out 1 and 3 months after the end of treatment.

RESULTSEA at Zhigou (TE 6) could obviously improve clinical symptoms and colon transmission time, decrease using rate of Kaisailu and cathartics, with an effective rate of 94.4%, which was better than 61.3% in the control group (P < 0.0001).

CONCLUSIONEA at Zhigou (TE 6) has a function of regulating qi and relaxing the bowels and it is an effective method for treatment of constipation with safety and no adverse effect.

Acupuncture Points ; Adult ; Aged ; Constipation ; therapy ; Electroacupuncture ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Qi

OBJECTIVETo search for an effective therapy for the acute stage of peripheral facial paralysis.

METHODSOne hundred and twenty cases of facial paralysis patients were randomly divided into an observation group and a control group, 60 cases in each group. Both groups were treated with routine medication, and the observation group was treated with shallow needling combined with acupoint application using self-made "acupoint application formula for facial paralysis" on the basis of the routine medication. The acupoints of Yifeng (TE 17), Wangu (GB 12), Dicang (ST 4), Jiache (ST 6), Yangbai (GB 14), Taiyang (EX-HN 5), Qianzheng (EX-HN 17), Cuanzhu (BL 2) in affected side and Hegu (LI 4) in healthy side were selected as main acupoints for shallow needling, and Qianzheng (EX-HN 17), Dicang (ST 4), Yangbai (GB 14) and Wangu (GB 12), or Jiache (ST 6), Taiyang (EX-HN 5), Quanliao (SI 18) and Yifeng (TE 17) were selected for acupoint application. The cure rate and the course of cured patients in both groups were compared.

RESULTSThe cure rate of 88.3% (53/60) in observation group was superior to that of 66.7% (40/60) in control group (P < 0.01). The course of cured patients in observation group was obviously shorter than that of control group (P < 0.01).

CONCLUSIONOn the basis of routine medication, shallow needling combined with acupoint application can improve the therapeutic effect and shorten the course for acute stage of peripheral facial paralysis.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Facial Paralysis ; therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the expression of CD34 in patients with acute promyelocytic leukemia (APL) and investigate the clinical and laboratory features of CD34(+) APL patients.

METHODS262 APL patients diagnosed by chromosome analysis and/or fusion gene examination in the last five years were retrospectively analyzed in this study. To survey the expression of CD34 in those patients, all the cases were divided into two groups (CD34(+) APL vs. CD34(-) APL). The clinical features including age, gender, abnormal values of the peripheral hemogram before treatment, the complete remission (CR) rate and the incidence of DIC and laboratory data such as the results of morphology, immunology, cytogenetics and molecular biology (MICM) between those two groups were compared.

RESULTSOf the 262 APL patients, 38 (14.5%) cases were positive for CD34 expression. There were no statistically significant differences between CD34(+) APL and CD34(-) APL groups in gender and age (P > 0.05). Before treatment, the median level of WBC in CD34(+) APL was 25.92 x 10(9)/L, which was significantly higher than that of CD34(-) APL (5.3 x 10(9)/L, P < 0.05). CD34(+) APL by morphology classification were mostly of the subtypes M3b and M3v (65.8%), while these subtypes in CD34(-) APL (40.3%) were significantly less (P < 0.01). There were no statistically significant differences between the two groups compared in respect of complete remission (CR) rate and the incidence of DIC (P > 0.05). The expression level of CD34 in APL had correlation to the expression level of CD2, CD7 and CD117; the latter three phenotypes in CD34(+) APL were significantly higher than those in CD34(-) APL (P < 0.01). No significant difference was found between those two groups by chromosome analysis, but there was more PML-RAR-alpha transcript short form in CD34(+) APL than that in CD34(-) APL (P < 0.05).

CONCLUSIONCD34(+) acute promyelocytic leukemia is a unique subtype of APL with different biological characteristics.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; blood ; Antigens, CD7 ; blood ; Antineoplastic Agents ; therapeutic use ; CD2 Antigens ; blood ; Child ; Disseminated Intravascular Coagulation ; etiology ; Female ; Humans ; Immunophenotyping ; Leukemia, Promyelocytic, Acute ; complications ; drug therapy ; genetics ; immunology ; Male ; Middle Aged ; Nuclear Proteins ; metabolism ; Phenotype ; Promyelocytic Leukemia Protein ; Proto-Oncogene Proteins c-kit ; blood ; Receptors, Retinoic Acid ; metabolism ; Remission Induction ; Retinoic Acid Receptor alpha ; Retrospective Studies ; Transcription Factors ; metabolism ; Translocation, Genetic ; Tretinoin ; therapeutic use ; Tumor Suppressor Proteins ; metabolism ; Young Adult

Adult ; Female ; Humans ; Hydrothorax ; diagnosis ; etiology ; therapy ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Talc ; therapeutic use ; Thoracoscopy ; methods