Objective To investigate the mechanisms by which nuclear factor-κB (NF-κB)translocates to nucleus of human glioblastoma U-87MG cells. Methods The levels of NF-κB translocating to nucleus in U-87MG after activation or inhibition ofphospholipase C-γ1 (PLCγ1) were first determined by electrophoretic mobility shift assay (EMSA), and then the levels of NF-κBtranslocating to nucleus in U-87MG after activation or inhibition of protein kinase-α (PKCα) were determined by EMSA. Results The level of NF- κB translocating to nucleus of U-87MG peaked 60min after EGF stimulation, which could be reversed by the pretreatment of PLCγ1 specific inhibitor U-73122. Moreover, PKC specific agonist phorbol myristate acetate (PMA) stimulation for 60 min promoted the nuclear translocation of NF-κB to the peak, which could be also effectively reversed by the pre-treatment of PKCα specific antagonist Ro 31-8220. Conclusions Epidermal growth factor can mediate the nuclear translocation of NF-κB in U-87MG through the signal transduction of PLCγ1-PKCα,thereby regulating the transcription of related invasion and metastasis genes.

In this study the chemical constituents of the higher polar sustances from Desmodium caudatum were investigated.The compounds were isolated by using column chromatographies over silicagel, polyamide, ODS, Sephadex LH-20, and preparative HPLC. The structures of these compounds were identified on the basis of NMR and MS spectra. Thirteen compounds were obtained and their structures were identified as vanillin(1), loliolide(2), indole-3-carboxaldehyde(3), salicylic acid(4), swertisin(5), saccharumoside C(6), isosinensin (7), kaempferol 3-O-β-D-glucopyranoside-7-O-α-L-rhamnopyranoside (8), isovitexin (9), vitexin (10), nothofagin(11), resveratroloside (12), and 2"-α-rhamnopyranosyl-7-O-methylvitexin (13). Except for compound 5, the remaining compounds were isolated from D. caudatum for the first time. Compounds 2, 3, 6-8, 11-13 were separated from the genus Desmodium for the first time.
Apigenin ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Fabaceae ; chemistry ; Molecular Structure ; Saponins ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization

Objective: To learn the results of MNA ( mini nutritional assessment ） nutrition screening and influencing factors in the elderly living at home, so as to provide basis for improving the nutritional status of the elderly living at home. Methods: The elderly people at home were recruited from Yinzhou District, Yiwu City and Changshan County in Zhejiang Province by the multi-stage random sampling method. Their demographic information, living habits and nutritional status were collected by the MNA scale and the questionnaire for nutrition and health status surveillance. The multivariate linear regression model was used to analyze influencing factors for the nutritional status. Results: Of 374 study subjects, 186 ( 49.73% ) were males and 188 ( 50.27% ) were females. The age was ( 69.63±6.68 ) years ( range, 60-90 years ). The average score of MNA scale was 25.26±2.81. The prevalence of malnutrition risk in the elderly living at home was 20.59%. Age ( β'=-0.140), marital status ( β'=0.110 ), annual income ( β'=0.155 ), active physical exercise ( β'= 0.104 ), eating health products/nutritional supplements ( β'= 0.110 ) and satiety ( full diet β'=0.196 ) were influencing factors for MNA scores ( P<0.05 ）. Conclusion The prevalence of malnutrition risk among the elderly living at home is 20.59%. The prevalence increases with age. Having a spouse, doing active physical exercise, eating health products/nutritional supplements, having healthy eating habits are conducive to maintaining the nutritional health of the elderly.

OBJECTIVETo investigate the preventive effects of a compound Danshen preparation (DSC) on long-term gastric lipid emulsion administration-induced nonalcoholic steatohepatitis in rats.

METHODSTwenty-seven 3-month-old SD rats were randomized equally into 3 groups and subjected to daily intragastric administration for 20 weeks of distilled water (control), lipid emulsion at 5 ml/kg (model group), and lipid emulsion plus DSC at 5.0 g/kg (DSC treatment group). After blood glucose (BG) determination, the rats were sacrificed for measurement of serum TC, TG, HDL-c, AST, and ALT, and the liver was weighed and pathologically examined.

RESULTSCompared with the control group, the rats in the model group showed significantly increased BG, TC, LDL-c, arteriosclerosis index (AI), AST, ALT, liver weight, and liver index (P<0.01) and decreased HDL-c (P<0.01), while TG remained unchanged. Fatty degeneration, hydropic degeneration and necrosis with inflammatory cell infiltration were observed in the liver of the rats in the model group. Compared with the model group, the rats in DSC groups showed decreased BG, AI (P<0.01), liver weight, liver index, AST, and ALT (P<0.05) and increased HDL-c, with milder pathological changes in the liver.

CONCLUSIONSLong-term gastric perfusion of lipid emulsion causes lipid metabolic disorder and nonalcoholic fatty liver disease in rats characterized by increased TC and decreased HDL-c. DSC can significantly increase HDL-c and provide partial protection of the liver against the damages by the lipid emulsion.

Animals ; Drug Administration Routes ; Drugs, Chinese Herbal ; therapeutic use ; Emulsions ; Fatty Liver ; chemically induced ; prevention & control ; Female ; Lipids ; administration & dosage ; toxicity ; Male ; Phenanthrolines ; therapeutic use ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Time Factors

OBJECTIVETo observe the effects of Danshen root compound (DSC) on blood lipid and bone biomechanics in mice with hyperlipemia-induced osteoporosis.

METHODSForty Kunming mice were randomized into 5 equal groups, and were given intragastric administration with distilled water (control), lipid emulsion (LE) at the daily dose of 5 ml/kg, LE plus simvastatin, LE plus DSC at 5.0 g/kg (DSC-L group), and LE plus DSC at 10.0 g/kg (DSC-H group), respectively. Serum TC, TG, and HDL-c levels and left femur hydroxyproline, calcium and phosphate contents were measured in the rats, with the right femur taken for bone biomechanical test.

RESULTSCompared with those in the control group, serum TC, LDL-c and AI of the mice increased and HDL-c, Hyp and bone calcium decreased significantly (P<0.01) with lowered bone biomechanical properties. Compared with those of the LE model group, AI decreased and HDL-c increased significantly in DSC-L and DSC-H groups (P<0.01), and the bone biomechanics in DSC-H group was improved.

CONCLUSIONLong-term intragastric administration of lipid emulsion causes lipid metabolic disorder and induces osteoporosis due to hyperlipemia in mice. DSC can significantly increase HDL-c and partially prevent the occurrence of osteoporosis in mice.

Animals ; Biomechanical Phenomena ; Bone and Bones ; drug effects ; metabolism ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Hyperlipidemias ; complications ; drug therapy ; Lipids ; blood ; Male ; Mice ; Osteoporosis ; blood ; etiology ; prevention & control ; Phenanthrolines ; pharmacology ; therapeutic use ; Phytotherapy ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo observe metabolomic changes in urine of chronic superficial gastritis (CSG) patients with Pi-qi deficiency syndrome (PQDS) or Pi-Wei dampness-heat syndrome (PWDHS), thereby providing scientific evidence for syndrome typing of them.

METHODSUrine samples were collected from CSG patients with PQDS/PWDHS and healthy volunteers, 10 in each group. Proton nuclear magnetic resonance spectroscopy (1H-NMR) based metabonomic analysis was performed on urine samples. Contents of related biomarkers were analyzed by principal component analysis (PCA), partial least square discriminant analysis (PLS-DA), and urivariate statistical analysis.

RESULTSPLS-DA analysis showed that metabolites among CSG patients with PQDS/PWDHS and healthy volunteers could be mutually distinguished. Seven differentially identified metabolites were screened from urines of CSG patients with PQDS and healthy volunteers included glutamate, methionine, α-oxoglutarate, dimethylglycine, creatinine, taurine, and glucose. Four differentially identified metabolites were screened from urines of CSG patients with PWDHS and healthy volunteers included 2-hydroxybutyric acid, trimethylamine oxide, taurine, and hippuric acid. Eleven differentially identified metabolites were screened from urines of CSG patients with PQDS and PWDHS included fucose, β-hydroxybutyric acid, alanine, glutamate, methionine, succinic acid, citric acid, creatinine, glucose, hippuric acid, and lactic acid.

CONCLUSIONThe metabolic differences of CSG patients PQDS and PWDHS mainly manifested in glycometabolism, lipid metabolism, and amino acids catabolism, and 1H-NMR based metabonomics may be used in classified study of Chinese medical syndrome typing.

Biomarkers ; urine ; Discriminant Analysis ; Gastritis ; urine ; Hot Temperature ; Humans ; Hydroxybutyrates ; Ketoglutaric Acids ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; physiology ; Metabolomics ; Principal Component Analysis ; Proton Magnetic Resonance Spectroscopy ; Qi ; Syndrome

This study was purposed to investigate the effects of siRNA targeting c-myc on apoptosis induction, proliferation in inhibition as well as c-myc protein and mRNA expression in human myeloid leukemia cell line HL-60 cells. C-myc siRNA synthesized in vitro was transfected into HL-60 cells by liposome. Changes of cell morphology were observed. Growth inhibition was detected by MTT assay and colony formation assay, and cell apoptosis was determined by DNA ladder. The expressions of c-myc mRNA and protein were detected by RT-PCR and Western-blot respectively. The results indicated that c-myc siRNA remarkably inhibited the cell proliferation, with an IC(50) value of 150 nmol/L. Data of DNA ladder showed that HL-60 cells apoptosis could be efficiently induced by c-myc siRNA, the apoptosis rate positively correlated with the time duration of treatment with drugs. The c-myc mRNA and protein expressions on HL-60 cells decreased after treatment with c-myc siRNA, which negatively correlated with time duration of treatment. It is concluded that c-myc siRNA can efficiently induce growth inhibition, decrease the expressions of c-myc mRNA and protein, and induce apoptosis in HL-60 cells.
Apoptosis ; genetics ; Cell Proliferation ; Gene Targeting ; HL-60 Cells ; Humans ; Leukemia, Myeloid ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo investigate the incidence of serum immunoglobulin (Ig) paraprotein in chronic lymphocytic leukemia (CLL), and to explore its clinical associated laboratory features and prognostic implication.

METHODSSerum protein electrophoresis and immunofixation electrophoresis were performed by automatic electrophoresis apparatus to identify serum Ig paraprotein. Immunonephelometry was used to measure serum Ig levels.

RESULTSOut of 101 CLL patients, serum Ig paraprotein detection was found in 20 (19.8%) cases, 13 (12.9%) patients with IgG paraprotein, 7 (6.9%) patients with IgM paraprotein and 1(1.0%) patient with IgA paraprotein. Among these 20 cases, 1 patient had both IgG and IgM paraprotein, 2 patients had both κ and λ light chains. The incidence of serum IgG paraprotein was high in the group of advanced Binet stage (P = 0.032) and high level of thymidine kinase 1 (TK1) (P = 0.013). The incidence of serum IgM paraprotein was high in the group of advanced Binet stage (P = 0.037), high level of TK1 (P = 0.017) and cytogenetic abnormalities of del(11q22.3) (P = 0.006). With a median follow-up of 30 months (range 1 - 101 months), 66 patients received therapy after initial diagnosis. Survival analysis showed that the patients with serum Ig paraprotein had significantly shorter treatment-free survival (TFS) times than the patients without serum Ig paraprotein (P = 0.024). And the patients with serum IgM paraprotein had significantly shorter treatment-free survival (TFS) times than the patients without serum Ig paraprotein (P = 0.013). However, serum Ig paraprotein or IgM paraprotein was not independent prognostic factor.

CONCLUSIONSerum Ig paraprotein can be detected in a subset of patients with CLL, which could be of value as a prognostic factor in CLL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Immunoglobulins ; blood ; Leukemia, Lymphocytic, Chronic, B-Cell ; blood ; diagnosis ; Male ; Middle Aged ; Paraproteins ; metabolism ; Prognosis ; Young Adult

OBJECTIVETo investigate the inhibitory effect of salidroside (Sal) on pulmonary microvascular endothelial cell (HPMEC) apoptosis induced by simulated microgravity and its mechanism.

METHODSHuman pulmonary microvascular endothelial cells cultured in vitro were divided into control group, clinorotation group and clinorotation+Sal pretreatment groups. Microgravity was simulated by clinorotation. The apoptotic rate of HPMECs was detected by flow cytometry using Annexin V-FITC staining, and the expressions of bcl-2, bax, and caspase-3 at the mRNA and protein levels were determined by real-time PCR and Western blotting, respectively.

RESULTSA 72-h clinorotation significantly induced apoptosis in HPMECs. Real-time PCR results demonstrated a significantly lowered bcl-2 but increased bax and caspase-3 mRNA expressions in clinorotation group as compared with those in the control group. Western blotting showed that clinorotation inhibited the protein expressions of PI3K and p-AKT and increased caspase-3 protein expression. Salidroside significantly inhibited the cell apoptosis, reversed the expressions of Bcl-2 and Bax, and attenuated the decrease in the protein expression of PI3K and phosphorylation level of AKT. Salidroside also antagonized the activation of caspase-3.

CONCLUSIONPI3K/AKT pathway and caspase 3 are involved in the apoptosis of HPMVECs induced by clinorotation, and the effect of clinorotation can be reversed by salidroside, suggesting the potential value of salidroside for application in spaceflight.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Endothelium, Vascular ; cytology ; drug effects ; Glucosides ; pharmacology ; Humans ; Lung ; blood supply ; Phenols ; pharmacology ; Signal Transduction ; Weightlessness

OBJECTIVETo investigate the effect of (AT)n repeat polymorphism of the 3'untranslated region in cytotoxic T-lymphocyte associated antigen 4 (CTLA-4) gene on CTLA-4 mRNA stability and full length (flCTLA-4) and soluble CTLA4 (sCTLA-4) expression in ulcerative colitis (UC).

METHODSflCTLA-4 mRNA in colonic biopsies and sCTLA-4 mRNA stability in peripheral blood mononuclear cells of UC patients were measured by quantitative PCR and half-life, respectively. The protein expression of flCTLA-4 in colonic biopsies and sCTLA-4 in sera of UC patients were determined by immunohistochemistry and enzyme-linked immunosorbent assay, respectively. The polymorphism of CTLA-4 (AT)n repeats in 300 UC and 700 age and sex matched healthy controls was genotyped by fluorescent PCR.

RESULTSAmong the UC patients, sCTLA-4 mRNA expression levels were decreased in active disease compared to non-active disease (P= 0.004). Carriers of the longer alleles of the (AT)n repeats expressed lower levels of flCTLA-4 and sCTLA-4 mRNA and sCTLA-4 protein than those of the shorter alleles in UC (all P< 0.01), and mRNA with long (AT)n repeat alleles has shorter half-life than mRNA with short alleles and, hence, are unstable. The frequency of long allele carriers of CTLA-4 (AT)n repeats was significantly higher in UC patients than in the healthy controls (22.0% vs. 6.3%, P< 0.01, OR= 4.21, 95% CI: 2.79-6.33), and associated with extensive colitis (P= 0.008).

CONCLUSIONCTLA-4 gene expression levels were associated with (AT)n repeat polymorphisms in UC patients. The expression of CTLA-4 mRNA and protein were decreased in carriers of the longer alleles of the (AT)n repeats of CTLA-4 gene. This study suggests that CTLA-4 plays an important role in genetic risk and pathophysiology for UC in central China.

Adult ; Antigens, CD ; chemistry ; genetics ; metabolism ; CTLA-4 Antigen ; Case-Control Studies ; Colitis, Ulcerative ; genetics ; metabolism ; Female ; Gene Expression Regulation ; Genotype ; Humans ; Male ; Phenotype ; Polymorphism, Genetic ; RNA Stability ; genetics ; RNA, Messenger ; chemistry ; genetics ; metabolism ; Solubility