Animals ; Calmodulin ; metabolism ; Epimedium ; Flavonoids ; pharmacology ; Gene Expression ; Hypothalamo-Hypophyseal System ; drug effects ; metabolism ; Male ; Pituitary-Adrenal System ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To develop an HPLC method for the measurement of n-3 fatty acid index of serum cholesteryl esters.Methods Serum triglycerides were hydrolyzed with ethanolic sodium hydroxide and cholesteryl esters (CEs) were extracted with hexane.The extracted CEs were analyzed by reversed phase HPLC with a UV detection at 205 nm.Cholesteryl eicosapentaenoate and docosahexaenoate ( major n-3 fatty acid cholesteryl esters) were identified by liquid chromatography-tandem mass spectrometry and cholesterol in each CE fraction was measured.Peak areas of CEs were corrected for cholesterol and CE n-3 index was calculated using the corrected peak area and expressed as the percentage of n-3 fatty acid CEs in total CEs.Results The HPLC analysis can be finished in 6 minutes.Triglycerides which interfere with the determination of n-3 fatty acid index, were hydrolyzed with ethanolic sodium hydroxide (4 mol/L) in 30 seconds.The within-run and total CVs for CE n-3 index averaged 0.66% and 0.90%, respectively.CE n-3 indexes of 70 volunteers and 36 coronary heart disease patients apparently healthy subjects and patients with coronary heart disease in Beijing Hospital appeared to be positively skewed and leptokurtic distribution ( skewness = 1.25, kurtosis = 1.70 ).The median of n-3 indices were 0.98% ( 0.37% - 2.40% ).The logarithm of n-3 index appeared to be normal distribution and the average is 0.003 7% with standard deviations of 0.15.The distribution of n-3 indices of gender groups was similar with the total.The medians of females and males were 1.08% (0.60% -2.40%) and 0.95% (0.37% -2.11%) respectively, and the former were significantly higher than the latter( t = - 3.021, P = 0.003 ).Conclusion A new method for the measurement of n-3 index of serum cholesteryl esters by HPLC has been established.It is simple and precise and can be used in predicting cardiovascular diseases risks and monitoring dietary intake of n-3 fatty acids.

This study is to investigate the anti-tumor activities of a novel cyclophosphamide derivate 4, 6-diphenyl cyclophosphamide (9b) in vivo and in vitro, and its possible mechanism of action. The inhibitory effects of 9b on human hepatoma cell line HepG2, human breast carcinoma cell line MCF-7 and human myeloid leukemia cell line K562 were measured by MTT assay in vitro. Cell cycle distribution and apoptotic rate were evaluated by flow cytometry. To evaluate the anti-tumor effect of 9b in vivo, mouse model bearing inoculated H22 tumor was established. The results indicated that 9b could inhibit the proliferation of HepG2, MCF-7 and K562 cells in a dose and time dependent manner. The ICo50 values of 9b were 32.34 micromol.L-1 to HepG2 cells, 87.07 micromol.L-1 to MCF-7 cells and 149.10 micromol.L-1 to K562 cells after incubation for 48 h. The results of flow cytometry indicated that after being treated for 48 h with different concentrations of 9b, the ratios of HepG2, MCF-7 cells at the Go/G1 phase and K562 cells at the G0/Gl phase and G2/M phase increased significantly compared with control group, and the apoptotic rate increased with the increase of the concentration of 9b. 9b could significantly reduce tumor weight of H22 solid tumor mouse model in vivo. To summarize, 9b showed significantly anti-tumor activity in vivo and in vitro, of which the mechanism might be associated with the change of cell cycle distribution and induction of tumor cell apoptosis.
Animals ; Antineoplastic Agents, Alkylating ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclophosphamide ; analogs & derivatives ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Humans ; Inhibitory Concentration 50 ; Liver Neoplasms, Experimental ; pathology ; Male ; Mice ; Molecular Structure ; Random Allocation ; Tumor Burden ; drug effects

0.05).In simple obesity group,there was positive correlation between BMI and urinary 24 h-Alb content(r=0.626,P

Objective To investigate the effects of partial body weight supported treadmill training (BW- STT) on post-stroke depression (PSD) and on patients' quality of life.Methods Sixty patients with PSD were re- cruited and divided into a training group (n=30,male 17,female 13) and a control group (n=30,male 16,fe- male 14).All patients were treated with routine internal medication and rehabilitation.The patients of the training group also received BWSTT in addition to their routine treatment.All patients' neurological impairment was evaluated using the Modified Edinburgh-Scandinavian Stroke Scale (MESSS).The Hamilton depression scale (HAMD) was used for evaluating the degree of depression.The Fugl-Meyer scale and the Barthel index were used to assess ambula- tion and balance,and facility in the activities of daily living.All patients were assessed before and after the treat- ment.Results After four weeks of treatment,depression in the training group had improved significantly more than in the control group.Conclusion BWSTT intervention is very important for patients with PSD:it can reduce the degree of depression and improve the quality of life.

Objective To observe the levels of alanine aminotransferase(ALT), total bilirubin(TBIL), hyaluronic acid (HA), procollagen type Ⅲ aminoterminal peptide (P Ⅲ NP) and larninin (LN) in the sera of rats infected with Clonorchis sinensis (C. sinensis) after treatment of albendazole combined with milkvetch root. Methods Thirty-two healthy adult Wistar rats were randomly divided into four groups with 8 in each based on body mass: control group, non-treatment group, Albendazole group(ALB group) and albendazole combined with milkvetch root group(ALB+MR group). The rats in non-treatmen, ALB and ALB+MR groups were infected orally with metacercariae of C. sinensis 50 per rat. The rats in control group were mock-infected with saline. The rats in ALB group were treated to each rat with 50 mg/kg alhendazole for 5 days, and ALB+MR groups were given to same treat with albendazole, meanwhile each rat injected with 800 mg/kg milkvetch root intraperitoneally for 30 days. All rats were killed after infestation 14 weeks and their sera samples were collected to detect ALT, TBIL, HA, PⅢNP, LN. Results There were statistically significant differences in the levels of ALT and LN in the sera of rats between groups(F=31.40,11.82, P<0.01). Compared with control[ (47.88±4.88)U/L, (51.20±4.12)μg/L], the levels of ALT and LN in rats in non-treatment group [(85.50±9.65)U/L, (64.20±4.18) μg/L] and ALB group [(65.29± 7.78) U/L, (58.23±2.55) μg/L] were significantly increased (P<0.05). Compared with non-treatment group, the levels of ALT and LN in rats in ALB group and ALB+MR groups[(50.25±9.29)U/L, (53.68±5.63)μg/L] were significantly decreased(P<0.05), and they decreased more obviously in ALB+MR group (P<0.05). There were statistically significant differences in the levels of TBIL, PⅢNP and HA in the sera of rats between groups (χ2=15.309,21.418,19.759, P<0.01). Compared with control[(0.700±0.350)μmol/L, (26.085±4.075)μg/L, (81.935±42.550)μg/L], the levels of TBIL, PⅢNP and HA in rats in non-treatment group(2.400 μmol/L, 46.220 μg/L,310.885 μg/L) and ALT group(1.200 μmol/L,36.540 μg/L, 178.010 μg/L) were significantly increased(P<0.05). Compared with non-treatment group, the level of TBIL in ALT+MR group(0.750 μmol/L), the levels of pⅢNP and HA in ALT and ALT+MR group(30.470,100.240 μg/L) were significantly decreased(P< 0.05). The levels of TBIL, PⅢNP and HA decreased more obviously in ALB+MR group(P<0.05). Conclusions The liver function in rats infected with C. sinensis is abnormal. The liver function and fibrosis are improved after treatment with albendazole or albendazole combined with milkvetch root. The treatment of albendazole combined with milkvetch root is more effective.

Objective To investigate the effect of excessive fluoride,aluminum on Zn,Fe,Ca,Mg,Cuin rat blood.Methods Forty eight SD rats were randomly divided into 4 groups matched with their weights:control group,high aluminum group,high fluorine group and high fluorine-aluminum group.Aluminum content in their drinking water was 0,90,0,90 mg/L respectively.Fluorine content of their feed was 5.2,5.2,106.0,106.0 mg/kg and aluminum Was 6.8,6.8,19.7,19.7 mg/kg respectively.90 days later,the level of blood Zn,Fe,Ca,Mg, Cu Was detected by the atomic absorption spectrometry.Results Compared among these groups,Zn,Fe,Mg and Cu content of the whole blood had significant difierences(F=46.25,14.74,6.10,2.93,P<0.05),while Ca content of the whole blood did not significantly change(F=2.81.P>0.05).Factorial analysis showed that excessive intake of aluminum could significantly decreased Zn,Fe,Mg content of the blood(F=42.66,5.41,7.04,P<0.05)and excessive intake of fluorine could significantly decreased Zn,Fe,Mg,Cu content of the blood(F=64.50,37.90,9.75,6.74, P<0.05).The coexistence offluorine and Muminum had interaction to the level of Zn(F=31.59,P<0.05)and did not obviously interact with other elements(F=0.91,1.63,1.51.0.00,P>0.05).Compared with the control group [(131.30 ±13.86)μmol/L,(10.24 ±1.02),(1.71 ±0.19)mmol/L,(20.43 ±4.42)μmol/L],Zn content in the high aluminum group[(90.84±9.98)μmol/L]decreased significantly(P<0.05),so did Zn,Fe,Mg content in tlle high fluorine group[(85.85 ±10.92)μmol/L,(8.49 ±0.68),(1.52 ±0.13)mmol/L],the difference being statistically significant(P<0.05)0Zn,Fe,Mg,Cu content in the high fluorine-aluminum group,being(82.82 ±11.00)μmol/L, (8.16±0.45),(1.46±0.09)mmoL/L,(15.69±2.38)μmol/L,respectively,all decreased signitlcarIdy(P<0.05). Compared with the high aluminum group[(9.43±1.09)mmol/L],Fe content of the high fluorine aluminum group[(8.16±0.45)mmol/L]decreased significantly(P<0.05).Conclusions Excessive fluoride can cause blood zn, Fe,Mg,Cu decline,so can excessive aluminum.Combination of excessive fluofine and aiuminum has 8ignificant synergic effect on the level of Zn but have rio influence on Ca.

The prenatal ethanol exposure induced the alterations of dendritic spine and synapse in visual cortex and their long-term effect would be investigated in mice from P0 to P30. Pregnant mice were intubated ethanol daily from E5 through the pup's birth to establish mode of prenatal alcohol abuse. The dendritic spines of pyramidal cells in visual cortex of pups were labeled with DiI diolistic assay, and the synaptic ultrastructure was observed under transmission electron microscope. Prenatal alcohol exposure was associated with a significant decrease in the number of dendritic spines of pyramidal neurons in the visual cortex and an increase in their mean length; ultrastructural changes were also observed, with decreased numbers of synaptic vesicles, narrowing of the synaptic cleft and thickening of the postsynaptic density compared to controls. Prenatal alcohol exposure is associated with long-term changes in dendritic spines and synaptic ultrastructure. The changes were dose-dependent with long term effect even at postnatal 30.
Animals ; Dendritic Spines ; ultrastructure ; Ethanol ; toxicity ; Female ; Fetal Alcohol Spectrum Disorders ; etiology ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal ; Microscopy, Electron, Transmission ; Pregnancy ; Prenatal Exposure Delayed Effects ; pathology ; Pyramidal Cells ; ultrastructure ; Synapses ; ultrastructure ; Visual Cortex ; ultrastructure

BACKGROUNDUsing hepatitis B virus e antigen (HBeAg) gene to construct the DNA-binding domain vector, which can express HBeAg in yeast cell, and can be used in yeast double hybrid as "bait plasmid" to look for the gene from the cDNA library, which expresses the protein that can interact with HBeAg.

METHODSPCR was performed to amplify the HBeAg gene from a sera of hepatitis B patient. The product of the amplification was inserted into T-vector and was verified by sequencing. Then it was inserted into the "bait" plasmid pGBKT7 after the digestion with the restricted endonuclease of EcoR I and Sal I. The plasmid was transformed into the yeast cell. PCR was used to verify whether the plasmid was transformed into yeast. The HBeAg protein expressed in the cell was confirmed by Western blot. Using nutrition selection assay to verify the constructed plasmid alone could not activate the reporter gene in the yeast cell.

RESULTSSequenced and digested by two endonucleases, the recombined vectors pGBKT7-eAg produced anticipated fragment. PCR verified that there was HBeAg fragment in the yeast. Having assayed by Western blotting, it was shown that the yeast cell transformed with pGBKT7-eAg vector had positive signal which could not be seen in the control. Tested by the nutrition selection assay, the recombined vectors pGBKT7-eAg could not activate LacZ reporter gene in the yeast.

CONCLUSIONDNA-binding domain plasmid was successfully constructed and could express HBeAg proteins in the yeast cell but could not activate transcription of LacZ reporter gene alone. The recombined plasmid can be used in yeast double hybrid.

Genetic Vectors ; Hepatitis B e Antigens ; biosynthesis ; genetics ; Hepatitis B, Chronic ; blood ; Humans ; Male ; Middle Aged ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; Two-Hybrid System Techniques ; Yeasts ; genetics

BACKGROUNDQuantitatively assessing myocardial perfusion and its reserve is of great importance for the diagnosis and stratification of patients with coronary artery disease (CAD), and represents an important goal of myocardial contrast echocardiography. In this study we sought to test the usefulness of low dose dobutamine stress real-time myocardial contrast echocardiography (RT-MCE) in the assessment of CAD, and to explore the relationship between perfusion reserve and contractile reserve.

METHODSTwenty-six patients with suspected or clinical diagnosed CAD were enrolled and underwent RT-MCE at baseline and under low dose dobutamine stress, and subsequent coronary angiography. RT-MCE images were analyzed quantitatively from microbubble replenishment curves for myocardial perfusion and its reserve.

RESULTSAt baseline, significant differences in beta (0.28 +/- 0.12, 0.25 +/- 0.09, 0.22 +/- 0.06, 0.20 +/- 0.07 respectively, P < 0.01) and A x beta (1.37 +/- 0.46, 1.28 +/- 0.47, 1.13 +/- 0.37, 0.91 +/- 0.32, respectively, P < 0.01) were observed among four segment groups with graded coronary artery stenosis severity (normal; 30% - 69% stenosis; 70% - 90% stenosis; and beyond 90% stenosis), but not observed in parameter A. When under stress, significant differences in A (5.73 +/- 1.28, 5.63 +/- 1.01, 4.96 +/- 0.81, 4.57 +/- 0.62, respectively, P < 0.01), beta (0.67 +/- 0.17, 0.55 +/- 0.19, 0.32 +/- 0.13, 0.25 +/- 0.08, respectively, P < 0.01) and A x beta (3.81 +/- 1.20, 3.11 +/- 1.17, 1.59 +/- 0.82, 1.12 +/- 0.37, respectively, P < 0.01) were observed among the formerly mentioned groups. Graded decreases in A reserve (1.20 +/- 0.53, 1.11 +/- 0.16, 0.98 +/- 0.12, 0.99 +/- 0.13, respectively, P < 0.01), beta reserve (2.65 +/- 1.07, 2.32 +/- 0.82, 1.44 +/- 0.40, 1.29 +/- 0.34, respectively, P < 0.01) and A x beta reserve (3.05 +/- 1.63, 2.59 +/- 1.01, 1.42 +/- 0.44, 1.27 +/- 0.34, respectively, P < 0.01) could also be observed with increasing coronary stenosis severity. In five segments groups scored by WMS (1 - 5), concordance between contractile function and myocardial perfusion could be found both at rest (beta: 0.28 +/- 0.11, 0.22 +/- 0.08, 0.21 +/- 0.05, 0.17 +/- 0.05, 0.19 +/- 0.06, respectively, P < 0.01; A x beta: 1.29 +/- 0.48, 0.98 +/- 0.45, 0.94 +/- 0.29, 0.76 +/- 0.30, 0.92 +/- 0.32, respectively, P < 0.01) and under stress (beta: 0.59 +/- 0.20, 0.35 +/- 0.15, 0.27 +/- 0.08, 0.17 +/- 0.05, 0.20 +/- 0.05, respectively, P < 0.01; A x beta: 3.07 +/- 1.38, 1.62 +/- 0.82, 1.28 +/- 0.40, 0.78 +/- 0.24, 0.93 +/- 0.22, respectively, P < 0.01). This concordance is also valid in terms of the reserves, and the MCE parameters in segments with ameliorated contractile function are significantly higher than in those without.

CONCLUSIONSQuantitative RT-MCE in conjunction with dobutamine stress shows promise in identifying and stratifying CAD and in exploring the perfusion-contractile correlation.

Adult ; Aged ; Contrast Media ; Coronary Angiography ; Coronary Circulation ; Coronary Disease ; diagnostic imaging ; physiopathology ; Dobutamine ; Echocardiography ; methods ; Female ; Humans ; Male ; Middle Aged ; Myocardial Contraction ; Reproducibility of Results