OBJECTIVETo study the effects of Dipsacus total saponins on the ability of learning and memory and its mechanism of action.

METHODSForty rats were randomly divided into blank control group, model group, Dipsacus group and positive control group (n = 10), general situation of rats were observed, the ability of learning and memory of rats was tested by Square water maze, the activities of acetylcholinesterase (AChE)and choline acetyltransferase (ChAT) of hippocampus in rats were measured using double antibody sandwich method.

RESULTSDuring the period of treatment, general situation had no obvious change in model group, but general situation and the ability of activity were gradually improved in Dipsacus group and positive control group. Compared with blank control group, the swimming time was obviously prolonged and the number of mistakes was obviously increased at different time, the activity of AChE was significantly enhanced and the activity of ChAT was significantly decreased in model group. Compared with model group, the swimming time was obviously shortened and the number of mistakes was obviously reduced at different time, the activities of AChE were significantly decreased and the activities of ChAT were significantly enhanced in Dipsacus group and positive control group; Compared with positive control group, the swimming time and the number of mistakes at different time and the activities of AChE and ChAT had no significant difference in Dipsacus group.

CONCLUSIONDipsacus total saponins can improve the ability of learning and memory in Alzheimer' s disease(AD) rats, its mechanism of 'action may be related to regulating ACh metabolism of hippocampus.

Acetylcholine ; metabolism ; Acetylcholinesterase ; metabolism ; Alzheimer Disease ; drug therapy ; physiopathology ; Animals ; Choline O-Acetyltransferase ; metabolism ; Dipsacaceae ; chemistry ; Disease Models, Animal ; Hippocampus ; drug effects ; Learning ; drug effects ; Memory ; drug effects ; Rats ; Saponins ; pharmacology

OBJECTIVETo Study the effect of anti-aging and its mechanism of total saponins of Wu-He Dipsacus asper on skin of mice-aging model.

METHODSForty-eight mice were randomly divided into blank control group, model group, low-Dipsacus group, medium-Dipsacus group, high-Dipsacus group and positive control group( n = 8) . The mouse model of skin aging was established by nape subcutaneous injection of 5% D-galactose (0.025 mL/(g · d)), the mouse of low-Dipsacus group, medium-Dipsacus group, high-Dipsacus group were administered with total saponins of Wu-He Dipsacus asper (50 ml/(kg · d), 100 mL/(kg · d), 200 mL/(kg · d)), the mice of the positive control group were administered with vitamin E(50 mg/(kg · d)) for 42 d. The content of hydroxyproline (HYP) and lipofuscin (LF) were measured in skin of each group mice, the activity of catalase (CAT) glutathione peroxidase ( GSH-Px) superoxide dismutase (SOD) and the content of malondi- aldehyde (MDA) were determined in serum and skin of each group mice.

RESULTSCompared with blank control group, the content of HYP decreased significantly and the content of LF increased significantly in skin, the activities of CAT, GSH-Px and SOD decreased significantly and the content of MDA increased significantly in serum and skin of model group; Compared with model group, the content of HYP increased significantly and the content of LF decreased significantly in skin, the activities of CAT, GSH-Px and SOD enhanced significantly and the con- tent of MDA decreased significantly in serum and skin of low-Dipsacus group, medium-Dipsacus group, high-Dipsacus group and positive control group; Compared with low-Dipsacus group, the content of HYP increased significantly and the content of LF decreased significantly in skin, the activities of CAT, GSH-Px and SOD enhanced significantly and the content of MDA decreased significantly in serum and skin of high-Dipsacus group and positive control group; The activity of SOD in serum and skin had a significant positive correlation with the content of HYP, and a significant negative correlation with LF in skin.

CONCLUSIONTotal saponins of Wu-He Dipsacus asper have obvious effect of anti-agng on skin of mouse-aging model , its mechanism is closely related to oxidative damage.

Animals ; Dipsacaceae ; chemistry ; Disease Models, Animal ; Mice ; Oxidative Stress ; Saponins ; pharmacology ; Skin Aging ; drug effects

OBJECTIVETo develop a method for determining epichlorohydrin in the workplace air by gas chromatograph-electron capture detector (GC-ECD).

METHODSEpichlorohydrin in the workplace air was collected by activated charcoal tubes, desorbed using acetone, and analyzed by GC-ECD.

RESULTSA good linearity was obtained in the range of 1.0-50 µg/mL (r=0.999 7). The detection limit was 0.012 µg/ml, while the recovery rate was 88.1% and relative standard deviation ranged from 1.11% to 3.57%. The samples could be stored for seven days at room temperature.

CONCLUSIONThis method effectively eliminates the interferences of alkanes on determination of epichlorohydrin and improves the sensitivity by 1 to 2 orders of magnitude, which can solve the problem of detection limit above standard in GBZ/T 160.58-2004.

Air Pollutants, Occupational ; analysis ; Charcoal ; Chromatography, Gas ; Epichlorohydrin ; analysis ; Workplace

Animals ; HIV Long Terminal Repeat ; genetics ; Humans ; Promoter Regions, Genetic ; genetics ; Spumavirus ; genetics ; Viral Proteins ; genetics ; metabolism

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) efficiently in pilot scale, cells of rhNDPK-A producing E. coli were homogenized by high pressure under 4 degrees C, 950 Pa. The insoluble debris was removed by microfiltration and the soluble portion was concentrated by ultrafiltration. The resulted crude sample was loaded on DEAE-sepharose Fast Flow. The target fraction was collected and then load on Cibacron Blue 3GA Sepharose CL-4B. Eluted with buffer containing ATP from the AC column, rhNDPK-A was polished with ultrafiltration. The results showed that after homogenized 2 rounds, 1500g cells of E. coli brought crude sample containing 47.6g NDPK-A. Treated with microfiltration and ultrafiltration, 27.3g of NDPK-A were recovered from this bacteria homogenate. After 2-step purification with column chromatography and then polished with ultrafiltration, 17.2 g rhNDPK-A were collected with purity of 96.3%. The recovery of the whole purification process was 36.2%, and the productivity of rhNDPK-A was 1.15 g per 100 g wet cells. Comparing the recovery of each purification step, it was found that the recovery of polish is higher than that of affinity chromatography, which is higher than that of ion exchange chromatography. The limit step was the process of sample pretreatment among the 4 purification steps. Combine with the fermentation results reported before, it was deduced that the productivity of rhNDPK-A was 510 mg/L. In conclusion, an easily controlled purification condition with high yield provides material for the translation researches of NDPK; In addition, it was suggested the crucial step determine the recovery of non-secretive recombinant proteins might be the process of sample pretreatment, not be the process of column chromatography.
Chromatography, Affinity ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Humans ; NM23 Nucleoside Diphosphate Kinases ; genetics ; metabolism ; Pilot Projects ; Recombinant Proteins ; isolation & purification ; metabolism ; Ultrafiltration

OBJECTIVETo study the function of IL-18 in promoting metastasis of lung cancer.

METHODSThe differential expression of IL-18 protein or mRNA level between highly and poorly metastatic sublines of human lung giant cell carcinoma metastatic model was detected by Western blot, semi-quantitative RT-PCR and northern blot analysis. The poorly metastatic PLA801C subline or highly metastatic PLA801D subline was transfected with constructed IL-18 sense or IL-18 antisense expressed plasmid by lipofectamine stable transfection technique. The metastasis-related effect mediated by IL-18, the metastatic phenotype differences, cell motility and cell invasion potential in vitro determined by MICS system and the expression level of metastasis-associated biomarkers detected by Western blot analysis, were compared between IL-18 stably transfectants and mock control, i.e. between PLA801C/IL-18(S) and PLA801C/pcDNA3.1, or between PLA801D/IL-18(As) and PLA801D/pcDNA3.

RESULTSIL-18 was only present in highly metastatic PLA801D subline at either protein or mRNA level, which implied that IL-18 might play a role in promoting metastasis of lung cancer. After IL-18 sense expressed plasmid was transfected into poorly metastatic PLA801C subline, IL-18 fused protein with myc tag detected by Western blot analysis using either IL-18 or myc tag monoclonal antibody. In addition, cell motility ability in vitro was significantly increased about 3 times and E-cadherin protein was significantly down-regulated at about 50% in PLA801C/IL-18(S) transfectants compared with mock control. While IL-18 expressed plasmid was transfected into highly metastatic PLA801D subline, IL-18 protein and mRNA were simultaneously decreased by 30%. In addition, cell invasion ability in vitro was significantly decreased at about 75% and E-cadherin protein was significantly up-regulated in PLA801D/IL-18(As) transfectants compared with mock control.

CONCLUSIONIL-18 might play a role in enhancing tumor metastasis of lung cancer by down-regulating E-cadherin protein expression.

Cadherins ; metabolism ; Carcinoma, Giant Cell ; metabolism ; secondary ; Cell Line, Tumor ; Cell Movement ; DNA, Antisense ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Interleukin-18 ; biosynthesis ; genetics ; Lung Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; genetics ; Plasmids ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

OBJECTIVETo study the hazards of gem dusts and the epidemiological features of the occurrence of gem worker's silicosis (GWS).

METHODSEpidemiological investigation in a gem factory in Guangdong province was carried out.

RESULTSThe average dusts concentration in workshop was 2.3 mg/m(3), average free silica content was 94.64%. There were 47 patients diagnosed as silicosis. The prevalence of GWS was 4.58% (47/ 1,027). The age of patients suffering from silicosis and the duration of dust exposure were (27.04 +/- 3.25) years and (4.67 +/- 1.17) years respectively. Chest radiographic examination showed mainly small round opacities "q". The incidences of GWS complicated by pulmonary tuberculosis, spontaneous pneumothorax and pulmonary emphysema were 10.64%, 10.64%, 44.68% respectively.

CONCLUSIONThe hazards of gem dust to workers might be serious. Higher incidence, younger age suffered from silicosis, shorter duration of dust exposure, more complications, and poorer prognosis in gem workers appeared to be the main epidemiological features of GWS in Guangdong.

Adult ; Air Pollutants, Occupational ; poisoning ; China ; epidemiology ; Female ; Humans ; Male ; Occupational Diseases ; epidemiology ; etiology ; Silicosis ; epidemiology ; etiology

OBJECTIVETo compare the clinical efficacy in the treatment of allergic rhinitis (AR) of lung qi deficiency and cold syndrome between Jin's three-needle therapy and western medication.

METHODSSixty-six patients were randomized into an acupuncture group and a western medication group, 33 cases in each one. In the acupuncture group, acupuncture was applied at three-nose points [Yingxiang (LI 20), Shangyingxiang (EX-HN 8) and Yintang (GV 29); Cuanzhu (BL 2) was added for frontal headache] and three-back points [Dazhu (BL 11), Fengmen (BL 12) and Feishu (BL 13)], once every day. Ten treatments made one session. Two sessions of treatment were required. In the western medication group, desloratadine oral suspension was prescribed, 5 mg each time, once a day, for 20 days. The scores of the symptoms and physical signs in AR patients as well as the clinical efficacy were observed between the two groups.

RESULTSThe total effective rate was 93.9% (31/33) in the acupuncture group, which was better than 72.7% (24/33) in the western medication group (P < 0.05). After treatment, the scores of AR symptoms and physical signs as well as the total score were all reduced compared with those before treatment in the two groups (all P < 0.01). The score of every item in the acupuncture group was lower than that in the western medication group after treatment (score of symptoms: 4.70 +/- 2.07 vs 6.55 +/- 2. 69, score of physical signs: 0.85 +/- 0.67 vs 1.45 +/- +0.62, total score: 5.36 +/- 2.70 vs 8.00 +/- 2.91, all P < 0.01).

CONCLUSIONJin's three-needle therapy achieves superior efficacy on AR of lung-qi deficiency and cold syndrome, which is better than desloratadine oral suspension.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adolescent ; Adult ; Child ; Female ; Humans ; Lung ; physiopathology ; Male ; Middle Aged ; Needles ; Qi ; Rhinitis, Allergic ; Rhinitis, Allergic, Perennial ; physiopathology ; therapy ; Young Adult

OBJECTIVETo investigate whether N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) can inhibit the differentiation of pulmonary fibroblasts into myofibroblasts by regulating Rho-associated coiled-coil forming protein kinase (ROCK) pathway mediated by transforming growth factor-β1 (TGF-β1).

METHODSPrimary culture of pulmonary fibroblasts was performed by trypsinization method. Four generations of pulmonary fibroblasts were divided into control group, TGF-β-induced differentiation group, Y-27632 treatment group, and Ac-SDKP treatment group. The intracellular distributions of ROCK, serum response factor (SRF), and α-smooth muscle actin (α-SMA) were observed by confocal laser scanning microscopy. The protein expression of ROCK, SFR, α-SMA, and type I and type III collagen in pulmonary fibroblasts was measured by Western blot. The mRNA expression of ROCK, SFR, and α-SMA was measured by real-time quantitative PCR.

RESULTSCompared with the control group, the pulmonary fibroblasts stimulated by TGF-β1 had a lot of α-SMA antibody-labeled myofilaments in parallel or cross arrangement, as observed by confocal laser scanning microscopy, and the mRNA and protein expression of ROCK, SRF, and α-SMA and protein expression of type I and type III collagen increased significantly after 6, 12, and 24 h of stimulation (P < 0.05). Compared with the TGF-β1-induced differentiation group, the Y-27632 treatment group and Ac-SDKP treatment group had significantly decreased mRNA and protein expression of ROCK, SRF, and α-SMA and protein expression of type I and type III collagen at the same time point (P < 0.05).

CONCLUSIONAc-SDKP can inhibit the differentiation of pulmonary fibroblasts into myofibroblasts and the synthesis of collagen in rats by regulating the ROCK pathway mediated by TGF-β1. That may be one of the mechanisms by which Ac-SDKP acts against (silicotic) pulmonary fibrosis.

Actins ; metabolism ; Animals ; Animals, Newborn ; Cell Differentiation ; drug effects ; Cells, Cultured ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Fibroblasts ; cytology ; drug effects ; Lung ; cytology ; drug effects ; Myofibroblasts ; cytology ; drug effects ; Oligopeptides ; pharmacology ; Rats ; Rats, Wistar ; Serum Response Factor ; metabolism ; Transforming Growth Factor beta ; pharmacology ; rho-Associated Kinases ; metabolism

OBJECTIVETo investigate the regulatory effect of N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP) on the activation of c-jun N-terminal kinase (JNK) signal transduction pathway and its role in silicotic fibrosis.

METHODSA rat model of silicosis was developed by intratracheal instillation. Sixty rats were randomly divided into 4-week control group (n = 10), 8-week control group (n = 10), 4-week silicosis model group (n = 10), 8-week silicosis model group (n = 10), AcSDKP treatment group (n = 10), and AcSDKP prevention group (n = 10). The content of hydroxyproline in lung tissue was measured using a p-dimethylaminoben-zaldehyde reagent; the expression levels of transforming growth factor (TGF)-beta 1 (TGF-β1), phospho-JNK, JNK, and c-jun in lung tissue were measured by Western blot. The lung fibroblasts from neonatal rats were cultured, and the 4th generation of cells were used in the experiment; these cells were divided into control group, TGF-β1 stimulation group, SP600125 intervention group, and AcSDKP intervention group. The distributions of phospho-JNK and c-jun in lung fibroblasts were observed by immunocytochemistry; the expression levels of type I collagen and type III collagen in lung fibroblasts were measured by Western blot.

RESULTSThe expression levels of TGF-β1, phospho-JNK, and c-jun and the content of hydroxyproline in the AcSDKP treatment group were 70.60%, 78.03%, 79.85%, and 71.28%, respectively, of those in the 4-week silicosis model group (P < 0.05) and 77.99%, 66.73%, 69.94%, and 64.82%, respectively, of those in the 8-week silicosis model group (P < 0.05); the expression levels of TGF-β1, phospho-JNK, and c-jun and the content of hydroxyproline in the AcSDKP prevention group were 84.56%, 61.18%, 64.73%, and 74.96%, respectively, of those in the 8-week silicosis model group (P < 0.05). The expression levels of phospho-JNK and c-jun in the AcSDKP intervention group were 54.59% and 55.56%, respectively, of those in the TGF-β1 stimulation group; the expression levels of type I collagen and type III collagen in the AcSDKP intervention group were 79.9% and 84.4%, respectively, of those in the TGF-β1 stimulation group (P < 0.05).

CONCLUSIONAcSDKP exerts anti-silicotic fibrosis effect probably by inhibiting the activation of JNK signal transduction pathway mediated by TGF-β1 and the deposition of interstitial collagen.

Animals ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; Male ; Oligopeptides ; pharmacology ; Pulmonary Fibrosis ; metabolism ; pathology ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Silicosis ; metabolism ; pathology