AIM:To investigate whether Smad pathway participates the process of extracellular signal regulated kinase (ERK) induced the proliferation of vascular smooth muscle cells (VSMCs). METHODS: Human umbilical artery smooth muscle cells (hUASMCs) were divided into four groups: control group, PDGF (platelet derived growth factor) group, ERK blocking agent group and PDGF+ERK blocking agent group. MTT assay was used to detect the proliferation of hUASMCs (A value). Immunohistochemical technique was used to detect the expression of PCNA, phosphorylated ERK (p-ERK) and phosphorylated Smad2/3 (p-Smad2/3) protein in hUASMCs. The expression of Smad2/3 mRNA in hUASMCs was detected by RT-PCR. RESULTS: The proliferation of hUASMCs and the expression of PCNA, p-ERK and p-Smad2/3 proteins in hUASMCs in PDGF group were increased obviously than those in other groups (P

Background and purpose:Ara-C is one of the most effective and common agents in the treatment of acute nonlyphocytic leukemia. Telomerase is a unique complex of ribonucleoprotein. It plays an important role in the pathogenesis and development of cancer. In this study, we investigate the changes of mRNA expression of telomerase subunits in HL-60 cells induced by Ara-c and try to come up with a theory that could help to assess the efficacy of Ara-C. Methods:The combinations of various Ara-C concentration and the incubation time were used to treat HL-60. The ratios of apoptotic cell to necrosis cell were determined by flow cytometry and the expressions of telomerase subunits mRNA were evaluated by RT-PCR.Results:① There was no influence on transcription of telomerase subunits gene after HL-60 cells was cultured with 0~0.2ug/ml Ara-C for 12 hours;② 2ug/ml and 10ug/ml of Ara-C could down regulate the expression of hTERT from 0.80+0.07 to 0.50+0.04 and 0.39+0.03, not hTR and hTP1;③ with longer incubation with 10ug/ml of Ara-C, the percentage of apoptosis could be increased. The maximal induction of apoptosis (18.16+4.25%) could be reached at 12hrs treatment of Ara-C, then gradually decreased later on. The rate of necrosis increased with time, the maximal percentage(57.94+12.03%) of necrosis was observed at 48hrs of incubation time with drug. The mRNA level of hTERT gene also decreased along with the cultured time , the lowest value (0.18+0.03) has been documented at 48hrs time point, but not hTR、TP1.Conclusions:① Ara-C could down-regulate the expression of hTERT mRNA in a dose-and time-dependent manner, but not hTR、hTP1;② There might be no relationship between the percentage of apoptosis induced by Ara-C apoptosis and the expression of telomerase hTERT gene mRNA, but a close relationship between necrosis and the expression of hTERT mRNA has been found.

2 years),seizure frequency(≥1 time/month),persistence time(≥60 s),gene-ralized seizure were all associated with the incidence of the loss of neuron in ammonias.Multivariate Logistic regression analysis showed that the independent influencital factors for the loss of neuron in ammonias in children with temporal epilepsy included seizure frequency,persis-tence time and tape of seizure. Conclusions The loss of neuron in ammonias though 1H-MRS can be detected.The results of multivariate analysis verify that the development of the loss of neuron in ammonias may be associated with many factors including age of onset,course of di-sease,seizure frequency,persistence time and generalized seizure.In order to lower the incidence of the loss of neuron,early intervening treatment is very important.

In order to find new cephalosporin with more and more potent antibacterial activity, nine new fourth-generation cephalosporins (N1-N9) were synthesized from ethyl 2-(2-aminothiazol-4-yl)-(Z)-2-methoxyiminoacetate (1) via acylation, substitution, hydrolysis, active esterification, condensation and salt formation. The structures of compounds (N1-N9) were confirmed by IR, MS, 1H NMR and elemental analysis. The target compounds possess different antimicrobial activities against Gram-positive and Gram-negative bacteria. The preliminary results of antibacterial activities revealed that they showed better antibacterial activities against Gram-positive bacteria than cefpirome sulfate. In particular, their activities against Staphylococcus aureus and Streptococcus albus are better.
Anti-Bacterial Agents ; chemical synthesis ; chemistry ; pharmacology ; Cephalosporins ; chemical synthesis ; chemistry ; pharmacology ; Gram-Negative Bacteria ; drug effects ; Gram-Positive Bacteria ; drug effects ; Molecular Structure ; Staphylococcus aureus ; drug effects ; Streptomyces ; drug effects

Objective To observe the effects of chloroquine phosphate on apoptosis of leukemic cell line U937, and investigate whether chloroquine phosphate induces leukemic cell apoptosis by normalizing protein PNAS-2's abnormal subcellular location. Methods Chloroquine phosphate of different concentrations were added into culture fluid of leukemic cell line U937 at logarithmic phase. MTr was used to measure cell proliferation, flow cytometry and laser confocal microscopy were applied to detect cell apoptosis, and immunofluorescence technology was employed to observe the effects of chloroquine phosphate on the changes of subcellular location of protein PNAS-2. Results Apoptosis of leukemic cell line U937 was significantly induced by 50 μg/mL chloroquine phosphate, and subcellular location of protein PNAS-2 was changed. Conclusion Chlorequine phosphate can induce apoptosis of leukemic cell line U937, and the mechanism may be related to the normalization of PNAS-2's abnormal subcellular location in U937 cell line. Chloroquine phosphate has the potential to be used in leukemic therapy.

Adult ; Ascorbic Acid ; administration & dosage ; analysis ; Humans ; Male ; Nutrition Surveys ; Occupational Health ; Riboflavin ; administration & dosage ; analysis ; Thiamine ; administration & dosage ; analysis ; Vitamins ; administration & dosage ; analysis

"Treatment for before sick" is a theory of TCM, reflecting preventing thought of "prevention being better than cure" and "nipping in the bud", while "moxibustion treatment for before sick" is highly praised by doctors of past ages. Moxibustion can activate human vital-qi and increase immunologic function of the organism, playing a preventive role for before sick, which is similar to the essence of "ischemic preconditioning" raised in recent years. Because of convenient manipulation, no adverse effect, it has very important position in the field of preventive medicine.
Acupuncture Points ; Humans ; Ischemic Preconditioning ; Moxibustion ; methods ; Preventive Medicine

OBJECTIVETo investigate the expression of LTF mRNA in several nasopharyngeal cancer (NPC) cell lines, and analyze the relationship between the genetic and epigenetic changes and expression of LTF gene.

METHODSThe expression level of LTF was detected in NPC cell lines HNE1, HNE2, HNE3, CNE1, CNE2, 5-8F, 6-10B cells and tissues of 15 cases of chronic nasopharyngitis by RT-PCR. The LTF protein level was analyzed by Western blotting in 6-10B cells. Then LOH, mutation and methylation status of LTF was examined by microsatellites analysis, PCR-SSCP, MSP and bisulfite genomic sequencing, respectively.

RESULTS15 chronic nasopharyngitis tissues showed stable LTF expression, while there were weak expression in 6-10B cells and absent expression in remaining detected NPC cell lines. There was a significantly lower LTF expression in chronic nasopharyngitis tissues (Z = -3.738, P = 0.000). No LTF protein expression was observed in 6-10B cells. LOH analysis demonstrated that allele loss of LTF wasn't found in NPC cell lines. LTF mutation was noted in 14.3% (1/7) of NPC cell lines. DNA sequencing confirmed the mutation point in the promoter region (-305 bp to -50 bp) was at -218 bp (del T) of LTF gene in the HNE1 cell line. Methylation of LTF gene was not found in chronic nasopharyngitis. However, methylation of LTF promoter was detected in all NPC cell lines. LTF mRNA expression was increased in 5-8F and 6-10B cell lines after treatment with 5-aza-2-deoxycytidine.

CONCLUSIONThere is an inactivation of expression of LTF gene in the NPC cell lines. Its molecular mechanism may be related with methylation of promoter region and deletion mutation.

Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Line, Tumor ; DNA Methylation ; Epigenesis, Genetic ; Gene Deletion ; Humans ; Lactoferrin ; genetics ; metabolism ; Loss of Heterozygosity ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; Nasopharyngitis ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; metabolism

Objective To investigate the influence of high intensity interval exercise (HIIT) on peroxidation and vascular endothelial function for experimental hyperhomocysteinemia (HHcy) rats. Methods Thirty five male rats were randomly divided into 4 groups. Control group (n=8) was given ordinary feed. High methionine group (n=27) was given 3% methionine on this basis, and divided into model group, folic acid group and HIIT+ folic acid group, with 9 rats per group for 16 weeks. Serum homocysteine (Hcy) , content of plasma malondialdehyde (MDA) , hydroxyl radical (OH-), total antioxidant capacity (T-AOC), activity of glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) were measured, as well as the level of Nitric Oxide (NO), Nitric Oxide Synthase (NOS) and Endothelin 1 (ET-1) . The pathology of abdominal aortas was analyzed.Results Sixteen weeks after intervention, there was no significant difference between HIIT + folic acid group and the control group (P>0.05) . The levels of serum Hcy in the model group, folic acid group and the HIIT+folic acid group were (23.95±3.35) μmol/L,(8.73±0.60) μmol/L, and (6.19±0.34) μmol/L respectively (P<0.05) . Sixteen weeks after intervention, the content of MDA in HIIT+ folic acid group reduced, and there was no significant difference compared with the control group (P>0.05). The level of SOD and GSH-PX increased in HIIT+ folic acid group and folic acid group, and there was a significant difference compared with the model group. There were significant differences in activities of SOD and GSH-PX in HIIT+ folic acid group when compared with folic acid group (P<0.05). Compared with the control group, there were significant differences in levels of ET-1, NOS and NO in folic acid group (P<0.05), while there was no significant difference in the level of ET-1 and NOS between HIIT+folic acid group and control group (P>0.05) . Mild atherosclerotic lesions were observed in the HIIT+folic group. Conclusion High methionine diet can reduce the level of serum Hcy in HHcy rats, and high intensity interval exercise combined with folic acid intervention could reduce the level of serum Hcy, improve oxidative stress state, reduce the injury of endothelial function, and thus to alleviate atherosclerotic lesion.

Solid dispersions technique was used to solidify buagafuran and improve buagafuran in vitro dissolution and stability. Buagafuran solid dispersions were prepared separately using PVPK30, PEG6000 and Poloxamer188 at various weight ratios as carriers. The status of buagafuran in solid dispersions was determined by using DSC and IR. The solubility, content and in vitro dissolution of pure drug and the solid dispersions were detected by using HPLC. When buagafuran/carrier was 1:5 or less, the drug existed in a solid dispersion form. Three kinds of carriers all can improve buagafuran dispersibility and in vitro dissolution. Accelerating experiment showed that buagafuran/PVPK30 < or = 1:10 solid dispersions was ageing-resistant, and the aspect, content and in vitro dissolution did not change after storaged over 3 months, but PEG6000, Poloxamer188 and a lower ratio PVPK30 solid dispersions became aged. Buagafuran/PVPK30 < or = 1:10 solid dispersions can be developed as buagafuran oral drug delivery carrier.
Anti-Anxiety Agents ; administration & dosage ; chemistry ; Drug Carriers ; Drug Delivery Systems ; Drug Stability ; Drug Storage ; Poloxamer ; chemistry ; Polyethylene Glycols ; chemistry ; Povidone ; chemistry ; Powders ; Sesquiterpenes ; administration & dosage ; chemistry ; Solubility