Objective To evaluate the effects of a health education project on endemic fluorosis in Shandong Province,and to provide a basis for formulating control strategies.Methods From December 2010 to June 2011,according to historical conditions,a total of 19 counties (cities,districts) of Shandong Province were chosen,and 3 townships (towns) were chosen in each project county.Health educational activities on endemic fluorosis were carried out in the Central Primary School in grade 4 to 6 in each township(town).In each project township(town),3 villages were chosen in each selected township(town) where the health educational activities in the community were carried out.Before and after the health educational activities,surveys on knowledge questionnaire on drinking-water-borne fluorosis control were conducted among 30 students of grade 5 in the Central Primary School and 15 housewives in every school location in each selected township(town).Results After the health educational activities,the knowledge awareness rates of endemic fluorosis control of the students and housewives were 96.53％ (5482/5679) and 94.88％ (3501/3690),respectively,and increased significantly compared with those before intervention [62.31％ (5154/8271) and 76.91％ (2815/3660)],and the difference was statistically significant (x2 =2176.50,490.58,all P ＜ 0.01).Among the primary school students and housewives,the knowledge awareness rates of endemic fluorosis control were increased by 34.22％ and 17.97％,respectively.Conclusions Health education activities on endemic fluorosis can significantly improve the knowledge awareness of target population,which will play a positive role in promoting prevention and control of endemic fluorosis.

Objective To investigate the anti-apoptotic effect of gene A20 in treatment of trau-matic brain injury (TBI). Methods Thirty-five Sprague-Dawley rats were made severe TBI models and assigned randomly to experimental group and control group (35 rats in each group). After severe TBI, the rats in experimental group were injected with liposome-pcDNA3.1-A20 and those in control group injected with liposome pcDNA3.1-A20 at 30 minutes after severe TBI. The animals in both groups were sacrificed to remove the brain of five rats from each group at 12, 24, 48, 72 and 168 hours for sec-tioning. The expression of A20 and neurocyte apoptosis were defined by immunohistological method and TUNEL accordingly. The other ten rats were testified for neurological function at 1,2, 3 and 4 weeks af-ter TBI. Results The expression of A20 in experimental group was higher than that in control group, with statistical differences (P < 0. 01). The peak neurocyte apoptosis was found at 72 hours after TBI. The number of apoptosis cells in experimental group was lower than that in control group at 12, 24, 48 and 72 hours afte TBI (P < 0.01 or 0.05). At the 4th week after TBI, the neurological function in exper-imental group was better than that in control group (P < 0.05). Conclusion Gene therapy with A20 may have anti-apoptosis effect and exert neuroprotective effect on severe TBI.

OBJECTIVETo investigate the effects of perinatal thyroid hormone deficiency on the expression of androgen receptor (AR) mRNA in cerebral cortex and hippocampus of rats.

METHODSPerinatal hypothyroidism was induced by the administration of propylthiouracil (PTU) solution to the dams by gavage (50 mg/d) beginning at embryonic d15 throughout the lactational period. In the T(4) injected group hypothyroid rats were injected intraperitoneally with levothroxine (L-T(4)) 2 microg/100 g BW daily, starting from the day of birth. Cerebral cortex and hippocampus specimen were collected from controls,hypothyroid and T(4)-injected hypothyroid rats on postnatal d1, 5, 10, 15 and 20. Quantification of ARmRNA in cerebral cortex and hippocampus was performed with competitive RT-PCR using internal and external standardization.

RESULTAge-related increasing ARmRNA levels were observed in neonatal rats, and those in male animals were significantly higher. AR expression was higher in the hippocampus than in the cerebral cortex. ARmRNA levels in the hypothyroid pups were lower than those in age-matched controls. The mRNA levels in the T(4)-injected hypothyroid pups were significantly higher compared with the age-matched hypothyroid pups, but in hippocampus ARmRNA expression did not reach normal levels in male rats at d10 and d20, in female at d15 and d20.

CONCLUSIONThe expression of ARmRNA decreases in brain of rats with perinatal hypothyroidism. Treatment with thyroid hormone can recover ARmRNA expression in cerebral cortex, but not in hippocampus.

Animals ; Animals, Newborn ; Cerebral Cortex ; metabolism ; Female ; Hippocampus ; metabolism ; Hypothyroidism ; metabolism ; Pregnancy ; Pregnancy Complications ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Receptors, Androgen ; biosynthesis ; genetics

OBJECTIVETo observe the effects of subchronic exposure to benzo[a]pyrene (B[a]P) on the mRNA and protein expression levels of apoptosis-related genes (bax, bcl-2, caspase-3, caspase-6, and caspase-9) and the activities of Caspase-3, Caspase-6, and Caspase-9 in the hippocampal neurons of rats and to investigate the neurotoxic mechanism by which B[a]P induces the apoptosis of neurons.

METHODSFifty-two healthy SD rat were randomly divided into five groups according to preliminary neurobehavioral test results: blank control group, solvent control group, and 1.0, 2.5, and 6.25 mg/kg B[a]P exposure groups; the rats in exposure groups were intraperitoneally injected with B[a]P every other day for 90 days. The Morris water maze was used to test the learning and memory ability of rats; flow cytometry was used to measure the apoptosis ratio of hippocampal neurons; real-time quantitative PCR and Western blot were used to measure the mRNA and protein expression levels of apoptosis-related genes; spectrophotometry was used to measure the activities of their en-coded proteins.

RESULTSCompared with the blank control group, solvent control group, and 1.0 mg/kg B[a]P exposure group, the 2.5 and 6.25 mg/kg B[a]P exposure groups hada significantly longer mean escape latency period (P < 0.05) and a significantly increased number of times of platform crossing (P < 0.05), and the 6.25 mg/kg B[a]P exposure group had significantly lower length and percentage of time spent in the platform quadrant (P < 0.05). The early apoptosis ratio rose as the dose of B[a]P increased (P trend < 0.05); the early apoptosis ratios of 1.0, 2.5, and 6.25 mg/kg B[a]P exposure groups were significantly higher than those of blank control group and solvent control group (P < 0.05). Compared with the blank control group, solvent control group, and 1.0 and 2.5 mg/kg B[a]P exposure groups, the 6.25 mg/kg B[a]P exposure group had significantly increased Bax expression (P < 0.05) and significantly decreased Bcl-2 expression and Bcl-2/Bax ratio (P < 0.05). The 2.5 and 6.25 mg/kg B[a]P exposure groups had significantly higher expression levels of Caspase-3 and Caspase-6 than the blank control group, solvent control group, and 1.0 mg/kg B[a]P exposure group (P < 0.05). The activities of Caspase-3, Caspase-6, and Caspase-9 were significantly higher in the 2.5 and 6.25 mg/kg B[a]P exposure groups than in the blank control group and solvent control group (P < 0.05). There was a positive correlation between the activities of Caspase-3, Caspase-6, and Caspase-9 and early apoptosis ratio of hippocampal neurons in rats (r = 0.793, P = 0.019; r = 0.886, P = 0.006; r = 0.773, P = 0.025). There were no significant differences in the mRNA expression of Bax, Bcl-2, Caspase-3, Caspase-6, and Caspase-9 among these groups (P > 0.05).

CONCLUSIONSubchronic exposure to B[a]P can induce apoptosis of hippocampal neurons; its mechanism may be related to the fact that B[a]P can induce upregulated expression of Bax, inhibit expression of Bcl-2, lead to decrease in Bcl-2/Bax ratio, induce upregulated expression of Caspase-3 and Caspase-6, and cause increase in the activities of Caspase-3, Caspase-6, and Caspase-9.

Animals ; Apoptosis ; drug effects ; Benzo(a)pyrene ; toxicity ; Caspases ; metabolism ; Hippocampus ; cytology ; drug effects ; Male ; Neurons ; drug effects ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

In order to study the anti-viral mechanism of extracted ZG from Gardenia, the effect of extracted ZG on Hep-2 cell membrane potential, Na -K+-ATPase activity and membrane fluidity post infected with parainfluenza virus type 1 (PIV-1) was observed. Acetylcholine which was fluorescent labeled with DiBAC4 (3) was taken as positive control to observe the changes of membrane potential and was measured by flow cytometer. The phosphorus determination method and spectrophotometer were used to measure the Na+-K+-ATPase activity of Hep-2 cell membrane post PIV-1 infection. Hep-2 cell membrane phospholipids was labeled with fluorescent NBD-C6-HPC and membrane fluidity was measured by confocal laser scanning microscope. The results demonstated that after PIV-1 infection the Hep-2 cell membrane potential decreased significantly and the membrane was in the state of hyperpolarization, Na+-K+-ATPase activity increased and membrane fluidity decreased significantly. There was no apparent interferring effect of extracted ZG on the changes of membrane potential and Na+-K+-ATPase activity post PIV-1 infection, while membrane fluidity was improved significantly. Acetylcholine improved the state of hyperpolarization. The changes of membrane potential, Na -K+-ATPase activity and membrane fluidity might be the biomechanism of PIV-1 infectoin. The extracted ZG improved membrane fluidity to prevent from PIV-1 infection by protecting the cell membrane, which was probably the mechanism of anti-PIV-1 activity of the extracted ZG, but ZG probably had nothing to do with membrane potential and Na+-K+-ATPase activity.
Acetylcholine ; pharmacology ; Antiviral Agents ; pharmacology ; Cell Line, Tumor ; Cell Membrane ; drug effects ; Gardenia ; chemistry ; Humans ; Membrane Fluidity ; drug effects ; Membrane Potentials ; drug effects ; Parainfluenza Virus 1, Human ; drug effects ; Plant Extracts ; pharmacology ; Sodium-Potassium-Exchanging ATPase ; metabolism

AIMTo study the effects of ginkgolide B on lipopolysaccharide (LPS)--induced TNFalpha production in mouse peritoneal macrophages and NF-kappaB activation in rat pleural polymorphonuclear leukocytes.

METHODSL929 crystal violet staining assay was used to show the level of TNFalpha released from mouse peritoneal macrophages induced by LPS. Electrophoretic mobility shift assay (EMSA) was used to determine NF-kappaB binding activities.

RESULTSGinkgolide B (1, 10 micromol x L(-1)) was shown to significantly inhibit LPS (10 mg x L(-1))-induced TNFalpha production in mouse peritoneal macrophages, the IC50 was 0.26 micromol x L(-1); LPS (1 mg x L(-1)) and PAF (1 nmol , L(-1)) were shown to increase the NF-kappaB binding activities in rat pleural polymorphonuclear leukocytes; ginkgolide B (10 micromol x L(-1)) was found to inhibit LPS (1 mg x L(-1))-induced NF-kappaB activation in rat pleural polymorphonuclear leukocytes; ginkgolide B (1, 10 micromol x L(-1)) was shown to inhibit PAF (1 nmol x L(-1))-induced NF-kappaB activation in rat pleural polymorphonuclear leukocytes.

CONCLUSIONThe inhibition of NF-kappaB activation and TNFalpha production might be considered to be part of the mechanisms underlying the antiinflammatory action of ginkgolide B; PAF is involved in activation of the NF-kappaB pathway stimulated with LPS.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Diterpenes ; isolation & purification ; pharmacology ; Female ; Ginkgo biloba ; chemistry ; Ginkgolides ; Lactones ; isolation & purification ; pharmacology ; Lipopolysaccharides ; antagonists & inhibitors ; Macrophages, Peritoneal ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; metabolism ; Neutrophils ; enzymology ; Plants, Medicinal ; chemistry ; Platelet Activating Factor ; antagonists & inhibitors ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; biosynthesis

OBJECTIVETo provide a basis for development and preparation of the new anti-tumor agents from Cortex A-canthopanacis senticosus (CAS), through isolating the active substances from CAS and studying the anti-tumor effect of CAS extracts in vivo and in vitro.

METHODSThe effects of CAS extracts and its isolated ingredients on tumor cell proliferation in vitro was determined by 3H-TdR incorporation; the anti-tumor component of CAS was isolated and purified by chromatography; the tumor bearing mice model was established by injecting tumor cell subcutaneously, and the model was used to observe the anti-tumor effect of CAS extract administered through gastrogavage.

RESULTSCAS extract showed obvious inhibition on tumor cell proliferation originated from multiple tissues (P < 0.01) and displayed a better dose-effect relationship. After orally taken CAS extract, the general condition of mice in the experimental group were better than that in the untreated control group, revealing a slower growth and significantly prolonged survival period (P < 0.01). A protein component, having inhibitory effect on tumor cell proliferation and the molecular weight was 64 kda, it was isolated by the thin layer gel chromatography.

CONCLUSIONCAS has not only the in vitro inhibitory effect on cell proliferation of multiple kinds of tumor, but also a good anti-tumor effect in vivo. The anti-tumor activity of CAS is correlated with a protein component with the molecular weight of 64 kda. Further isolation, purification, study on mechanism will provide scientific evidence for clinical application and development of CAS in anti-tumor effect.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Eleutherococcus ; Female ; HL-60 Cells ; pathology ; Humans ; Leukemia, T-Cell ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Tumor Cells, Cultured

Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species (ROS) levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.

OBJECTIVETo study the antiproliferation effect on HepG2 cells and its underlying mechanism of the active chemical composition of the Viburnum Odoratissimum.

METHODS3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and trypan blue dye exclusion assay were used to assess the effect of vibsane-type diterpenoids on the proliferation of various tumor cells. Alterations in cell cycle and apoptosis were determined by flowcytometry. The enzymatic activity of caspase-3/7 was measured by Apo-ONE homogeneous Caspase-3/7 Assay kit.

RESULTSCompound 1 #, a vibsane-type diterpenoid, was found to significantly inhibit the growth of HepG2 cells by anticancer proliferation activity screening. It was demonstrated that the modified groups on side chain coupled to C11 site affected the cell growth-inhibition activity of compounds by structure-activity analysis. In addition, HepG2 cell line was most sensitive to compound 1 #, which induced growth arrest of HepG2 cells in a dose- and time-dependent manner. Study on the mechanisms underlying these effects indicated that compound 1 # induced significant G0/G1 phase arrest of HepG2 cells in a time- and concentration-dependent manner. Meanwhile, It was found that higher concentrations of compound (5-10 micromol/L) caused evident increase in the unmber of apoptotic cells and dose-dependent activation of caspase-3/7.

CONCLUSIONVibsane-type diterpenoids could significantly inhibit the growth of HCC HepG2 cells. Induction of cell cycle arrest and apoptosis may play important roles in their anticancer effects.

Apoptosis ; drug effects ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Diterpenes ; pharmacology ; Hep G2 Cells ; Humans ; Viburnum ; chemistry

This study is to explore the effects of extracts of Cheezheng pain relieving plaster (ECPRP) on nitric oxide (NO) production and expression of inducible nitric oxide synthase (iNOS) in macrophages induced by LPS and the mechanism involved. Nitric oxide level was measured with Griess reagent assay. Inducible nitric oxide synthase (iNOS) protein and NF-kappaBp65 fragment were detected with Western blotting. ECPRP (62.5 and 125 mgL(-1)) significantly inhibited the increase of nitric oxide level. Furthermore, ECPRP (62.5 and 125 mg x L(-1)) notably reduced the expression of iNOS mRNA and protein. ECPRP (62.5 and 125 mg x L(-1)) elevated the content of I-kappaB protein in cytoplasm, while decreased the content of NF-kappaBp65 protein in nucleus. These results suggest that ECPRP reduce nitric oxide level via down-regulation of NF-kappaB-iNOS-nitric oxide pathway, resulting in prevention of inflammation.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Lipopolysaccharides ; adverse effects ; Macrophages ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; RNA, Messenger ; genetics ; Transcription Factor RelA ; metabolism