Child ; Humans ; Plasma Exchange ; Thrombotic Microangiopathies ; therapy

OBJECTIVETo study the effect of astragalosides (AST) on the anoxia/reoxygenation (A/R) injured neuron in rat.

METHODSPrimary cultured rat's hippocampal neurons were made into A/R model cells. The cell viability was detected by MTT assay and lactate dehydrogenase releasing methods; the activity of superoxide dismutase (SOD), and contents of malondialdehyde (MDA) and nitride oxide (NO) in culture supernate were detected; the apoptosis rate of hippocampal neurons after A/R was measured by flow cytometry with double-staining of Hoechst33258 and AnnexinV-PI; and intracellular calcium ion [Ca2+]i was observed with a cofocal laser-scanning microscope and determined by fluorescent probe Fluo-3/AM.

RESULTSAST enhanced the cell viability of neurons after A/R injury, increased SOD activity and decreased the MDA and NO contents in supernate, reduced the A/R-induced apoptosis and decreased the calcium overload in neurons.

CONCLUSIONAST has the protective effects on A/R injured neurons, the mechanism is possibly related with its anti-oxidation and calcium overload reducing actions.

Animals ; Calcium ; metabolism ; Cell Hypoxia ; drug effects ; Female ; Fetus ; Hippocampus ; cytology ; Malondialdehyde ; metabolism ; Neurons ; cytology ; Neuroprotective Agents ; pharmacology ; Nitric Oxide ; metabolism ; Pregnancy ; Primary Cell Culture ; methods ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Saponins ; pharmacology ; Superoxide Dismutase ; metabolism ; Triterpenes ; pharmacology

Purpose:To compare the therapeutic and adverse effects between continuous and intermittent infusion of fluorouracil(5-FU) in advanced colorectal cancer.Methods:17 patients of advanced colorectal cancer were treated with continuous intravenous infusion of 5-FU 2.5g in 100 ml of 5% glucose in a continuous infusions pump,for 120 h (Group A).16 patients of similar severity were treated with 5-FU 500 mg in 500 ml of 5% glucose 2 h iv daily for 5 d (Group B). Total of 33 patients were treated with calcium folinate (CF) 200 mg in 100 ml of 5% glucose 1 h iv daily for 5 d and HCPT 10 mg in 250 ml of 5% glucose 2 h iv daily for 6d.The therapeutic and adverse effects were evaluated according to the cri- teria recommended by WHO.Results:Effectiveness was noted in 9 patients in Group A and 3 patients in group B.(x~2 test P

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 compare the regulatory effects of electro-acupuncture (EA) at acupoints Zusanli (ST36) and Hegu (LI4) on the visceral hyper-sensitivity in the rat model of irritable bowel syndrome (IBS), and to explore the acting targets and specialty of acupoints.

METHODSExcept 8 rats of the normal control group, the rest 32 rats were prepared to set up the IBS models. IBS animal model was prepared by enema with acetic acid. Model rats were divided into three groups. Except for rats in the model group for control, those in the other two groups were treated 20 min by EA on ST36 (EA-ST36) and LI4 (EA-LI4) respectively for 2 weeks to observe the effect on behavior response of viscera sensitivity. The changes of neuropeptide (NPY), the somatostatin (SS) levels in blood and tissues of brain and intestine were monitored as well.

RESULTSThe volume thresholds for abdomen uplifting and back hunching were obviously increased after EA-ST36 (P<0.05), but showed insignificant change after EA-LI4. NPY contents lowered and SS contents increased in model rats; both EA-ST36 and EA-LI4 could raise the level of thalamic NPY (P<0.01 and P<0.05, respectively), but showed insignificant effects on NPY in colonic tissue. As for SS content, its colonic level could be reduced by EA-S36 and EA-LI4 (P<0.01 and P<0.05, respectively), however, its blood level was affected only by EA-ST36 (P<0.05).

CONCLUSIONSEA-ST36 or EA-LI4 could regulate the NPY in thalamus and SS in colonic tissue, the former could affect blood level of SS as well. It is deemed that NPY and SS may be the key substances for regulating the action of acupuncture in the brain-intestinal axis; their different levels could be regarded as an indicator for the functional difference between the acupoints.

Acupuncture Points ; Animals ; Brain ; metabolism ; Electroacupuncture ; methods ; Irritable Bowel Syndrome ; metabolism ; physiopathology ; Neuropeptide Y ; metabolism ; Rats ; Rats, Wistar ; Somatostatin ; metabolism ; Viscera ; physiopathology

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

To produce specific monoclonal antibody (McAb) against human thrombomodulin (hTM), the full-length hTM cDNA-expressing plasmid pThr402 was transfected into CHO cells by Lipofectamine 2000 reagent. The hTM-expressing CHO cells, which was confirmed by flow cytometry and Western blot, were obtained by G418 selection. Then the McAb against hTM was prepared with classic hybridoma technique. A cell line of CHO-TM5 with high level of hTM was used to immunize female Balb/c mice 3 times at an interval of 4 weeks. On the third day after the third immunization, mice were sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96-well plates for screening. Cellular enzyme-linked immunoabsorbent assay (CELISA) was applied twice. The first CELISA was done with polythene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but with CHO cell monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5(+)CHO(-) hybridoma cells. Balb/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded McAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. One line of hybridoma cells with high expression of specific McAb against hTM, NH-1, was obtained. The Ig subclass of the McAb was IgG1 and the titer of ascitic McAb was 1x10(-6). Flow cytometry, CELISA and Western blot assays demonstrated that McAb NH-1 could specifically recognize hTM expressed in CHO-TM5 cells and human umbilical vascular endothelial cells. Meanwhile, the tissue specificity of antigen recognized by McAb NH-1 was identified by immunohistochemical ABC staining. NH-1 can specifically recognize the natural hTM expressed mainly in vascular endothelial cells, which will potentially be useful for investigation of the functions and clinic values of hTM.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; CHO Cells ; Cricetulus ; Female ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Thrombomodulin ; immunology ; Transfection

OBJECTIVETo study the effects of exogenous ER beta on the growth of breast cancer MCF-7 cells under different treatment.

METHODSAn eukaryotic expression vector containing 1.6 kb of human entire coding sequence of ER beta (pCDNA3-ER beta) was transfected into human breast cancer MCF-7 cells using lipofectamine 2000. The biological activity of ER beta was detected with the luciferase reporter containing estrogen responsive element (ERE) and the expression of ER beta protein by Western blot. The growth properties of MCF-7, pCDNA 3-transfected MCF-7 and pCDNA 3-ER beta-transfected MCF-7 cells under different treatment, including E2 (17beta-estradiol) and 4-OHT (4-hydroxytamoxifen), were observed.

RESULTSA stronger activation of the reporter by ER beta in the presence of E2 was observed in the pCDNA 3-ER beta-transfected MCF-7 cells than in the pCDNA 3-transfected MCF-7 and in MCF-7 cells. Western blot analysis showed that the protein level of ER beta in the pCDNA 3-ER beta-transfected MCF-7 cells was markedly increased. Exogenous ER beta expression did not change the growth properties and the morphology of MCF-7 cells under normal condition. The pCDNA 3-ER beta-transfected MCF-7 cells proliferated at the same rate as naive cells in the presence of 4-OHT, whereas a strong inhibition of the proliferation of the pCDNA 3-ER beta-transfected MCF-7 cells in the presence of E2 was observed.

CONCLUSIONExogenous ER beta expression does not increase the resistance to 4-OHT, and a strong inhibition of the proliferation may occur in the presence of E2.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Estradiol ; pharmacology ; Estrogen Antagonists ; pharmacology ; Estrogen Receptor beta ; genetics ; metabolism ; Female ; Humans ; Tamoxifen ; analogs & derivatives ; pharmacology ; Transfection

OBJECTIVETo investigate the expression of programmed death-1 (PD-1) and programmed death ligand-1(PD-L1) in peripheral T-lymphocytes from patients with chronic periodontitis and its significance and to clarify its role in the development of chronic periodontitis.

METHODSA total of 73 subjects were included in the study and divided into three groups, chronic periodontitis(30 cases), chronic gingivitis(25 cases) and 18 healthy controls. The peripheral blood was collected and PD-1/PD-L1 expression in the surface of CD(+)4 T lymphocytes and CD(+)8 T lymphocytes was examined by flow cytometry. Blood samples from 16 chronic periodontitis patients were collected at week 0 and 6 after initial therapy for 6 weeks and PD-1 and PD-L1 expression in the surface of CD(+)4 and CD(+)8 T lymphocytes was also determined by flow cytometry. The data were statistically analyzed.

RESULTSThe percentage of PD-1 expression in CD(+)4 and CD(+)8 T lymphocytes of chronic periodontitis group[(16.7 ± 5.5)%,(20.8 ± 5.1)%]and chronic gingivitis group[(14.2 ± 6.1)%,(14.5 ± 4.3)%]were higher than that of healthy controls[(9.5 ± 2.1)%, (8.1 ± 1.9)%](P < 0.05). The percentage of PD-L1 expression in CD(+)4 and CD(+)8 T lymphocytes of chronic periodontitis group[(24.2 ± 7.1)%,(15.3 ± 6.8)%]and chronic gingivitis group[(12.4 ± 6.0)%,(11.2 ± 5.5)%]were higher than that of healthy controls[(4.7 ± 1.2)%, (3.2 ± 2.3)%] (P < 0.05). The percentage of PD-1/PD-L1 expression in CD(+)4 T lymphocytes and CD(+)8 T lymphocytes of the chronic periodontitis group were significantly decreased after initial therapy(P < 0.05).

CONCLUSIONSThe expression of PD-1 and PD-L1 in peripheral CD(+)4 T lymphocytes and CD(+)8 T lymphocyte of chronic periodontitis patients was up-regulated and was associated with periodontal condition. The initial therapy reduced the expression of PD-1 and PD-L1.

B7-H1 Antigen ; biosynthesis ; Case-Control Studies ; Chronic Periodontitis ; immunology ; metabolism ; Flow Cytometry ; Humans ; Programmed Cell Death 1 Receptor ; biosynthesis ; T-Lymphocytes ; Up-Regulation

OBJECTIVETo observe the effects of two main isoflavones, daidzein and genistein on the bone-nodule formation in rat calvaria osteoblasts in vitro.

METHODSOsteoblasts obtained from newborn Sprague-dawley rat calvaria were cultured for several generations. The second generation cells were cultured in Minimum Essential Medium supplemented with ascorbic acid and Na-beta-glycerophosphate for several days, in the presence of daidzein and genistein, with or without the estrogen receptor antagonist ICI 182780. Number of nodules was counted at the end of the incubation period (day 20) by staining with Alizarin Red S calcium stain. The release of osteocalcin, as a marker of osteoblast activity, was also determined on day 7 and day 12 during the incubation period.

RESULTSCompared with the control, the numbers of nodules were both increased by incubation with daidzein and genistein. 17 beta-estradiol was used as a positive control and proved to be a more effective inducer of the increase in bone-nodules formation that daidzein and genistein. The release of osteocalcin into culture media was also increased in the presence of daidzein and genistein, as well as 17 beta-estradiol on day 7 and day 12 (day 12 were higher). The estrogen receptor antagonist ICI 182780 completely blocked the genistein- and 17 beta-estradiol-induced increase of nodule numbers and osteocalcin release in osteoblasts. However, the effects induced by daidzein could not be inhibited by ICI 182780.

CONCLUSIONThese findings suggest that geinistein can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts. The effects, like those induced by 17 beta-estradiol, are mediated by the estrogen receptor dependent pathway. Daidzein also can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts, but it is not, at least not merely, mediated by the estrogen receptor dependent pathway.

Animals ; Cells, Cultured ; Genistein ; pharmacology ; Isoflavones ; pharmacology ; Osteoblasts ; cytology ; drug effects ; metabolism ; Osteocalcin ; biosynthesis ; Osteogenesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Skull ; cytology ; drug effects ; metabolism