Animals ; Anticholesteremic Agents ; pharmacology ; Cholesterol ; blood ; Drug Evaluation, Preclinical ; methods ; Male ; Rats

Objective To establish a rat model of coal-burning-borne fluorosis,and to observe the expression changes of bone morphogenetic protein-2 (BMP-2) and bone morphogenetic protein-3 (BMP-3) in the serum of rat treated with different dose of fluoride and different treatment duration.Methods A total of 120 clean grade SD rats(body mass between 80 to 120 g) weaned for 4 weeks were randomly assigned into four groups,which were control,low-dose fluorid,medium-dose fluorid and high-dose fluorid groups,respectively,and 30 rats in each group (female 15,male 15).All of the rats were fed with coal drying corn from fluorosis area.Ten rats were killed by femoral artery bleeding 30 d,90 d and 180 d after exposed to fluoride,respectively.Serum BMP-2 and BMP-3 level was tested by enzyme-linked immunosorbent assay (ELISA).Results ①Results of BMP-2:after exposed to fluoride for 90 d and 180 d,the differences of serum BMP-2 level between groups were statistically significant(F=385.08,173.98,all P ＜ 0.01).In low-dose fluorid,medium-dose fluorid and high-dose fluorid groups,the expression of serum BMP-2 at 90 d[(18.80 ± 0.43),(22.22 ± 0.85),(25.14 ± 0.69)μg/L] and 180 d[(7.98 ± 0.68),(8.97 ± 0.78),(15.04 ± 0.89)μg/L] was higher than that of control group[(12.54 ± 1.29),(7.53 ± 0.97)μg/L,all P ＜ 0.05],and the level of BMP-2 increased with increasing dose of fluoride (all P ＜ 0.05).Within each group,the difference of serum BMP-2 was statistically significant(F =55.42,511.58,686.35,671.64,all P ＜ 0.01).The expression of BMP-2 in each group at 90 d [(12.54 ± 1.29),(18.80 ± 0.43),(22.22 ± 0.85),(25.14 ± 0.69)μg/L] was higher than that at 30 d[(11.75 ± 1.15),(11.42 ± 1.07),(11.38 ± 0.92),(11.15 ±1.03)μg/L,all P ＜ 0.05].The expression of BMP-2 in each group at 180 d[(7.53 ± 0.97),(7.98 ± 0.68),(8.97 ± 0.78),(15.04 ± 0.89) μg/L] was lower than that at 90 d.②Results of BMP-3:the difference between groups was not statistically significant at every experimental stage(F =0.7215,1.2951,0.0964,all P ＞ 0.05).Conclusions Longer excessive fluoride intake stimulates the expression of BMP-2 in rats,but with prolonged fluoride intake,the stimulation becomes weak.The effect of fluoride on BMP-3 is not as sensitive as that on BMP-2.

There are 250 species of Zanthoxylum (Rutaceae) in the world. This genus distributed in tropical and subtropical regions. Alkaloids are the major and representative ingredients in these plants including quinolines, isoquinolines, and amide alkaloids, with such biological activities as anti-tumor, anti-inflammatory, analgesic, anti-virus, anti-platelet aggregation, anti-bacteria and anti- oxidant. These species have been used for a long time to treat toothache, urinary and venereal diseases, lumbago and rheumatism. This review summarizes the chemical constituents and pharmacological activities from the Z. sppplants, in an effort to the systematic research and application of the alkaloids of this genus.
Alkaloids ; chemistry ; pharmacology ; Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Molecular Structure ; Zanthoxylum ; chemistry

Objective To observe the effects of extracellular signal-regulated kinase (ERK) 1/2 and protein kinase B (Akt) signal pathway in cholangiocarcinoma cells invasion and migration promoted by microRNA-21 (miR-21).Methods The experimental study was adopeted.QBC939 cholangiocarcinoma cells were cultured in vitro,through constructing and synthesizing unrelated sequence,miR-21 mimics and miR-21 inhibitor which were transfected into cells,and these cells were allocated into 4 groups,including growing naturally cells in the cell group,cells transfected by unrelated sequence in the 21-NC group,cells transfected by miR-21 mimics in the 21-M group and cells transfected by miR-21 inhibitor in the 21-Ⅰ group.Besides,cells in the 21-M group were allocated again into the 2 groups,20 μmol/L LY294002 and 10tμmol/L U0126 were respectively added in order to dispose 48 hours for follow-up experiments.Indicatiors of the test:(1) real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-21 in each group of cholangiocarcinoma cells.(2) Werstern blot was performed to detect the relative expressions of PTEN,ERK and Akt proteins in each group of cholangiocarcinoma cells.(3) Scarification assay was executed to test the migration of each group of cholangiocarcinoma cells.Transwell experiment was conducted to examine the migration and invasion of each group of cholangiocarcinoma cells.The measurement data with normal distribution were presented by x-s.The means of the 2 groups were compared by the t test.The means among groups were compared by the ANOVA,and pairwise comparison was analyzed by the Bonferroni test.The repeated measurement data were analyzed by the repeated measures ANOVA.Results (1) The relative expression of miR-21 in the cell group,21-NC group,21-M group and 21-Ⅰ group were 1.010 ±0.010,0.980 ± 0.050,4.900 ± 0.350 and 0.260 ± 0.010,respectively,with a statistically significant difference among the 4 groups (F =78.23,P ＜ 0.05),with no statistically significant difference between the 21-NC group and cell group (P ＞0.05).There was increased expression between the 21-M group and cell group,decreased expression between the 21-Ⅰ group and cell group and significant difference between 21-M group or 21-Ⅰ group and cell group (P ＜ 0.05).(2) The relative expressions of PTEN,ERK,p-ERK,Akt and p-Akt proteins in the cell group,21-NC group,21-M group and 21-Ⅰ group were 0.360 ± 0.020,0.400 ± 0.030,0.140 ± 0.010,0.680 ± 0.110 and 0.045 ± 0.126,0.470 ± 0.140,0.460 ± 0.060,0.440 ± 0.110 and 0.310 ± 0.020,0.380 ± 0.040,0.590 ± 0.060,0.160 ±0.010 and 0.400 ±0.010,0.390 ±0.080,0.410 ±0.090,0.380 ±0.070 and 0.440 ±0.110,0.510 ± 0.120,0.980 ± 0.150,0.190 ±0.010,respectively,showing statistically significant differences among the4 groups (F =10.23,12.78,18.11,P ＜ 0.05).There was no significant difference in the relative expressions of PTEN,ERK,p-ERK,Akt and p-Akt proteins between the cell group and 21-NC group (P ＞0.05).Compared with cell group,there was decreased PTEN expression and increased p-ERK and p-Akt expressions in the 21-M group,showing statistically significant differences (P ＜ 0.05).Compared with cell group,there was increased PTEN expression and decreased p-ERK and p-Akt expressions in the 21-Ⅰ group,showing statistically significant differences (P ＜ 0.05).(3) The change of migration rate of cells from 6 hours to 48 hours were from 12.0％ ± 3.0％ to 23.0％ ± 5.0％ in the cell group,from 21.0％ ± 4.0％ to 43.0％ ± 7.0％ in the 21-M group,from 6.0％ ±1.0％ to 18.0％ ±4.0％ in the miR-21 + LY294002 group and from 9.0％ ±2.0％ to 26.0％ ± 6.0％ in the miR-21 + U0126 group,respectively.The migration rate of cells in the 21-M group at each time point was higher than that in the cell group (F =16.23,P ＜0.05).The migration rate of cells in the miR-21 + LY294002 group and miR-21 + U0126 group were lower than that in the 21-M group (F =25.21,P ＜ 0.05),and there was the interaction effects between the change of migration rate of cells of the 3 groups and time,with a statistically significant difference (F =35.31,P ＜ 0.05).(4) The numbers of migration cells in the cell group,21-M group,miR-21 + LY294002 group and miR-21 + U0126 group were 198 ± 32,248 ± 39,187 ±23 and 174 ± 28,respectively,with a statistically significant difference among the 4 groups (F =8.48,P ＜ 0.05) and between the 21-M group and cell group (t =4.13,P ＜0.05).Compared with the 21-M group,the numbers of migration cells in the miR-21 + LY294002 group and miR-21 + U0126 group were decreased (F =21.98,P ＜0.05).The numbers of invasion cells in the cell group,21-M group,miR-21 + LY294002 group and miR-21 + U0126 group were 102 ± 22,211 ± 36,55 ± 9 and 67 ± 13,respectively,showing a statistically significant difference among the 4 groups (F =11.32,P ＜ 0.05) and between the 21-M group and cell group (t =6.67,P ＜ 0.05).Compared with the 21-M group,the numbers of invasion cells in the miR-21 + LY294002 group and miR-21 + U0126 group were decreased (F =36.23,P ＜ 0.05).Conclusion ERK and Akt signal pathway participate in the cholangiocarcinoma cells invasion and migration promoted by miR-21,PTEN could mediate the process of promoting cholangiocarcinoma cells invasion and migration through ERK and Akt signal pathway promoted by miR-21.

BACKGROUND: It is difficult to cover aneurysm-like ventricular septal defect (VSD) of large inlet and multiple outlets completely with symmetrical type occluders or eccentric type occluders. OBJECTIVE: To investigate the feasibility of A4B2 occluder devices for covering aneurysm-like VSD, and to observe the effects of proper occhiders selected according to pseudoaneurysm size on coveting aneurysm-like VSD. DESIGN: Case analysis.SETTING: the First Hospital of Hebei Medical University. PARTICIPANTS: From August 2004 to May 2006, 226 patients with the pseudoaneurysm of petimembranous VSD, who underwent interventional therapy in the First Hospital of Hehei Medical University, were recruited in the study. According to the results of the left ventricular angiography, 36 patients of pseudoaneurysm of petimembranous VSD with large inlet and multiple outlets were closured with A4B2 occluder devices. According to the results of the left ventricular angiography, the mean diameter of the left inlet of VSD was (10.6+8.7) mm (ranged from 8 to 21 mm), the mean diameter of the right outlet of VSD was (3.1 ± 2.9) mm (ranged from 2 to 8 ram). Main materials: Occluder device and delivery mechanism were offered by Shanghai Shape Memory Alloy Materials Company and Beijing Starway Medical Technology Inc. They were processed into double disks using nickel-titanium shape memory alloy wires by a special technology to close VSD by a transcatheter approach. The size of the occluder was denoted with the diameter of the waist, and the Size ranged from 4 to 16 mm in the present study. METHODS: All the occluders were transferred by a 7-10 F transferring sheath from right heart system, and the mean diameter of the occluders was (6.364-2.48) nun (ranged from 4 to 16 ram). Fifteen minutes after the procedure, left ventricular angiography and transthoracic echocardiography (TIE) were performed again to evaluate the efficacy. After the procedure, electrocardiogram (ECG) monitoring lasted for 5 successive days in all patients, and ECG and TIE were performed 1, 3, 6 and 12 months later. MAIN OUTCOME MEASURES: Residual shunt, arrhythmia and valve function as well as blood compatibility. RESULTS: Sixteen cases were closured by placing the occhiders into left inlet of VSD, 16 cases were closured by placing the occluders into the pseudoaneurysm completely, and 4 cases were closured at the outlet of the defects. The results of the left ventricular angiography and TTE that performed fifteen minutes after the procedure demonstrated that 32 cases were completely closured and slightly residual shunts (< 3 mm) was found in other 4 patients. And confirmed by TIE, the residual shunts completely disappeared in 2 of the 3 patients 24 hours later while in the other one in 1 month after the procedure. Temporary left bundle branch block was found in 3 cases while temporary right bundle branch block was found in 2 cases, and all of them recovered within one week. Without severe complications, all of the 36 patients were treated successfully with A4B2 (thin waist shape) occluder devices made in China. Critical appraisal in blood compatibility of the implantation materials used in this research had been performed. The hemolysis ratio was less than 5%, the platelet adhesion was less, and the blood coagulation function ,the immune system response( immunoglobulin and complement)and the re-endothelialization of material surface were all normal. CONCLUSION: Transcatheter interventional therapy with domestic A4B2 occluder devices for VSD with pseudoaneurysm is safe, effective, promising, and has fewer complications. The key to the procedure is to select suitable occluders and suitable positions where to plant them according to the size, morphologic characteristics, position, and maturity of the pseudoaneurysm.

AIMTo develop a method for determination of loganin in mouse plasma by using high-performance liquid chromatography. The method was employed to study pharmacokinetics of loganin.

METHODSAn RP-C18 was used as the stationary phase. The mobile phase consisted of methanol-water (30:70), at the flow-rate of 0.8 mL.min-1. The UV absorbance detector was set at 240 nm. Plasma samples were treated with solid phase extraction.

RESULTSThe recovery of loganin in mouse plasma was 86.0%-91.5%. The calibration curve in plasma was linear over the range of 0.01-5.00 micrograms.mL-1. The limit of quantitation was 10 ng.mL-1. The RSDs of intra-day and inter-day (n = 5) were less than 15%. The pharmacokinetic parameters were Cmax = 6.8 micrograms.mL-1, Tmax = 30 min, T1/2 alpha = 26.1 min, T1/2 beta = 29.01 min.

CONCLUSIONThe method is accurate, sensitive and suitable for pharmcokinetic study of loganin. The absorption and elimination of loganin were rapid after ig in mice.

Adjuvants, Immunologic ; blood ; pharmacokinetics ; Animals ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Iridoids ; blood ; pharmacokinetics ; Male ; Mice

Objective：To evaluate bioequivalence and relative bioavailability of domestic and imported repaglinide tablets in healthy volunteers. Methods： Twenty two healthy male volunteers were randomized into A and B groups. A single dose (4 mg) of domestic and imported repaglinide tablets were given respectively according to an open 2-way crossover study design. The washout period was 1 week. Plasma concentrations of repaglinide were determined by HPLC method. Results：The pharmacokinetic parameters of domestic and imported drugs were as follows: t1/2 were(0.86±0.24) and (0.83±0.31) h；tmax were( 0.79±0.37) and (0.75±0.41) h；cmax were (52.43±20.92) and (53.32±24.94) μg/L. AUC0-t were (79.87±36.48) and (74.95±30.57) μg*h*L-1，respectively. The relative bioavailability of domestic formulation was (106.55±16.15)%. Conclusion： The results of variance analysis and two one-side t test show that 2 formulations are of bioequivalence.

OBJECTIVETo investigate the gene mutation and the molecular pathogenesis of an inherited coagulation factor VII (F VII) deficiency pedigree with consanguineous marriage.

METHODSThe diagnosis was validated by coagulant parameter assay on the prothrombin time (PT), activated partial thromboplastin time, fibrinogen and coagulation factor activity. F VII gene mutations were analyzed in the proband and other family members by direct DNA sequencing of the PCR products of all exons, exon-intron boundaries and 5'and 3' untranslated sequences. The mutations were confirmed by reverse sequencing.

RESULTSThe values of PT and F VII activity in the proband were significantly abnormal, they were 30.9 s and 3% respectively. The PT of her daughter, father and mother was slightly extended to 21.2 s, 16.3 s and 16.1 s respectively, and the F VII activity was reduced to 22%, 25% and 35% respectively. The coagulant parameters of her younger brother were within normal range. Homozygous T-->G transition at position 11482 in exon 8 was identified in the proband resulting in His348Gln, and heterozygosity for His348Gln was confirmed in her daughter and her parents, and the normal wild-type was observed in her younger brother.

CONCLUSIONHomozygous missense mutation of His348Gln was found in a pedigree of hereditary F VII deficiency. The mutation was inherited from her heterozygote parents.

Adolescent ; Adult ; Aged ; Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Female ; Homozygote ; Humans ; Infant, Newborn ; Male ; Middle Aged ; Mutation, Missense ; Pedigree

Aged ; Anti-Inflammatory Agents ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Female ; Humans ; Immunosuppressive Agents ; therapeutic use ; Methylprednisolone ; therapeutic use ; POEMS Syndrome ; complications ; diagnosis ; drug therapy ; pathology ; Plasma Cells ; pathology ; gamma-Globulins ; therapeutic use

BACKGROUNDAdoptive transfer of allogeneic tumor-specific T cells often results in severe graft-versus-host disease (GVHD). Here, we sought to maximize graft-versus-tumor and minimize GVHD by using haploidentical T cells in pre-irradiated B16-melanoma bearing mice.

METHODSC57BL/6 mice bearing B16-melanoma tumors were irradiated with 0, 5, or 7 Gy total body irradiation (TBI), or 7 Gy TBI plus bone marrow transplantation. Tumor areas were measured every 3 days to assess the influence of irradiation treatment on tumor regression. B16-melanoma bearing mice were irradiated with 7 Gy TBI; sera and spleens were harvested at days 1, 3, 5, 7, 9, 11, and 13 after irradiation. White blood cell levels were measured and transforming growth factor β1 (TGF-b1) and interleukin 10 (IL-10) levels in serum were detected using enzyme-linked immunosorbent assay (ELISA) kits. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry were performed to test TGF-b1, IL-10 and Foxp3 mRNA levels and the proportion of CD4+CD25+Foxp3+ T-regulatory cells (Tregs) in spleens. B16-melanoma bearing C57BL/6 mice were irradiated with 7 Gy TBI followed by syngeneic (Syn1/Syn2) or haploidentical (Hap1/Hap2), dendritic cell-induced cytotoxic T lymphocytes (DC-CTLs) treatment, tumor areas and system GVHD were observed every 3 days. Mice were killed 21 days after the DC-CTLs adoptive transfer; histologic analyses of eyes, skin, liver, lungs, and intestine were then performed.

RESULTSIrradiation with 7 Gy TBI on the B16-melanoma-bearing mice did not influence tumor regression compared to the control group; however, it down-regulated the proportion of Tregs in spleens and the TGF-b1 and IL-10 levels in sera and spleens, suggesting inhibition of autoimmunity and intervention of tumor microenvironment. Adoptive transfer of haploidentical DC-CTLs significantly inhibited B16-melanoma growth. GVHD assessment and histology analysis showed no significant difference among the groups.

CONCLUSIONAdoptive transfer of haploidentical tumor-specific T cells in irradiation-pretreated B16-melanoma bearing mice preserved antitumor capacity without causing a GVHD response.

Animals ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Graft vs Host Disease ; Immunotherapy, Adoptive ; methods ; Male ; Melanoma, Experimental ; metabolism ; therapy ; Mice ; Mice, Inbred C57BL ; Real-Time Polymerase Chain Reaction ; T-Lymphocytes ; immunology ; T-Lymphocytes, Regulatory ; immunology