Objective To observe the effects of MPP+ on the expression of inhibin, activin and its receptor ⅡA and cell ability in PC12 cells of rats.Methods The changes of expression of activin?A, activin?B, inhibin? and receptorⅡA mRNAs were assayed by RT-PCR, the change of cell viability was detect by tyrpan blue exclusion method in cultured PC12 cells at 3 h,6 h,12 h,24 h after MPP+ was added and compared with control groups.Results The expression of activin?A, activin?B and activin receptorⅡA mRNAs in PC12 cells were down-regulated significantly after MPP+ administrated at every time point while the expression of inhibin? mRNA remained unchanged. The cell viability decreased at the points of 12 h and 24 h after MPP+ administrated.Conclusion MPP+ may cause the injury of PC12 cells by downregulating the expression of activin and its receptor.

Objective To investigate the utility of diffusion weighted imaging (DWI) and blood oxygen level-dependent (BOLD) magnetic resonance imaging (MRI) in the assessment of renal hypoxia in an experimental model of mice with lupus nephritis (LN).Methods MRL/lpr mice (n=13) were studied and C57BL/6 mice (n=10) served as controls.Urinary albumin to creatinine ratio (ACR),serum creatinine (Scr),anti-ds-DNA antibody,and complement C3 levels were measured.The mice underwent coronal echo-planar DWI and BOLD MRI of the kidneys when they were 14-16 weeks old.Hypoxyprobe was administered intraperitoneally to the mice 1 hour before they were sacrificed.The distribution of HypoxyprobeTM-1,hypoxia-inducible factor 1 α (HIF-1 o) and heme oxygenase-1 (HO-1) in renal tissues was detected by immunohistochemical analysis and Western blotting.Results Urinary ACR,Scr and anti-ds-DNA antibody levels in MRL/lpr mice were significantly higher than that in C57BL/6 mice.It was found that HypoxyprobeTM-1,HIF-1o and HO-1 distributed widely in the renal tissue of MRL/lpr mice,and closely associated with the renal tubulointerstitial lesion.The mean apparent diffusion coefficient (ADC) value of kidneys in MRL/lpr mice was (1.52±0.27) × 10-3 mm2/s,and the mean R2* values of the renal cortex and medulla were (30.95 ±4.59)/s and (23.43± 3.06)/s respectively,all significantly lower than that in C57BL/6 mice (P=0.037,P=0.030 and P=0.043,respectively).The ADC of medulla was negatively correlated with urinary albumin to creatinine ratio (r=-0.364,P=0.032;r=-0.329,P=0.050),the ADC of cortex was negatively correlated with the level of serum creatinine (r=-0.814,P=0.014;r=-0.755,P=0.031) when b value was 500 s/mm2 and 800 s/mm2,and the mean R2* value was negatively correlated with the degree of tubulointerstitial lesions and the expression of hypoxia parameters (all P ＜ 0.05).Conclusions Renal hypoxia may play an important role in renal tubulointerstitial lesion.Functional MRI may be used to monitor renal function changes,pathological injuries and renal hypoxia in LN.

Cinobufacino injection is a significant anti-tumor medicine for the treatment of various tumors in clinic, which was made from water extraction of the skin of Bufo bufo gargarizans. In present paper, HPLC-DAD-FT-ICR-MS method was used to identify the major bufadienolides in cinobufacino for the first time. Solid-phase extraction with dichloromethane and silica was used to enrich the total bufadienolides in cinobufacino. Based on the UV and high resolution MS/MS data, 33 bufadienolides were analyzed and characterized. Among them, eight compounds were identified by comparing with standard references unambiguously. This study elucidated the major bufadienolides in cinobufacino, which provided material foundation of cinobufacino and will be benefit for the further pharmacological research.
Amphibian Venoms ; chemistry ; Animals ; Bufanolides ; analysis ; chemistry ; Bufo bufo ; Chromatography, High Pressure Liquid ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

OBJECTIVETo investigate the differential plasma protein profiles in patients with hyperlipidemia & atherosclerosis (H&A) of different patterns of phlegm-stasis syndrome (PSS) for seeking their biomarker proteins.

METHODSTwo-dimensional gel electrophoresis and gel screening graphical analysis were performed on plasma proteins got from 146 patients; corresponding protein spots were fetched from the gel for two-stage mass-spectrometric analysis by quadruple time-of-flight mass spectrometry; then the differential proteins for PSS were discriminated by Fisher discriminate analysis.

RESULTSExcepting two uncertain proteins, 7 differential proteins were screened out from the 11 differentially expressed plasma protein spots with variability over 100% in the inter-block matching. Classic analysis found that haptoglobin precursor and fibrinogen gamma chain were possibly the plasma biomarker proteins for H & A; fibrinogen beta chain and apolipoprotein A-I precursor were that set apart PSS from non-PSS; fibrinogen gamma chain, albumin and apolipoprotein A-I precursor were for phlegm syndrome; haptoglobin precursor, adrenomedullin binding protein precursor, albumin and complement component C4 were for stasis syndrome; albumin and adrenomedullin binding protein precursor were for the phlegm-stasis mutual blocking syndrome. Moreover, the above mentioned expressions of possible marker proteins had their own special rule of changing in the transforming progress of PSS.

CONCLUSIONThis study reported, for the first time, the existence of evident variation of functional protein constitution in different patterns of PSS, and definite compatibility being detected in some functional proteins, which may be the marker proteins for making diagnosis and prognosis of PSS in H&A. Besides, preliminary proof for the transformation of PSS has gained at the functional protein level.

Adult ; Atherosclerosis ; blood ; diagnosis ; Blood Proteins ; analysis ; Case-Control Studies ; Gene Expression Profiling ; Humans ; Hyperlipidemias ; blood ; diagnosis ; Medicine, Chinese Traditional ; Middle Aged ; Proteomics

Suitable pretreatment of biological samples can truly reflect the role of law of the measured components played in the body and will provide experimental evidence for the studies on metabolic process, material basis of efficacy, mechanism of action, pharmacology, toxicology and the others. Biological samples include blood, urine, hair, tears, etc. There are also many samples processing methods, such as the direct protein precipitation, liquid-liquid extraction and solid phase extraction and so on. These methods could be used alone or combined.
Animals ; Body Fluids ; chemistry ; Chemical Precipitation ; Chemistry Techniques, Analytical ; methods ; Humans ; Liquid-Liquid Extraction ; Proteins ; isolation & purification ; Solid Phase Extraction

The aim of this study is to prepare hyaluronic acid (HA) modified core-shell liponanoparticles (pHA-LCS-NPs) as gene delivery system and investigate its gene transfection efficiency in human retinal pigment epithelium (ARPE-19) cells in vitro. The pHA-LCS-NPs was prepared by firstly hydrating dry lipid film with CS-NPs suspension to get LCS-NPs, then modifying the lipid bilayer with HA by amidation reaction between HA and dioleoyl phosphatidylethanolamine (DOPE). Its morphology, particle size and zeta potential were investigated. XTT assay was used to evaluate the cell safety of different vectors in vitro. The gene transfection efficiency of pHA-LCS-NPs modified with different contents of HA was investigated in ARPE-19 cells with green fluorescent protein (pEGFP) as the reporter gene. The results showed that the obtained pHA-LCS-NPs exhibited a clear core-shell structure with the average particles size of (214.9 +/- 7.2) nm and zeta potential of (-35 +/- 3.7) mV. The 24 h cumulative release of gene from pHA-LCS-NPs was less than 30%. After 48 h incubation, gene transfection efficiency of pHA-LCS-NPs/pEGFP was 1.81 times and 3.75 times higher than that of CS-NPs/pEGFP and naked pEGFP, respectively. Also no obvious cytotoxicity was observed on pHA-LCS-NPs. It suggested that the pHA-LCS-NPs might be promising non-viral gene delivery systems with high efficiency and low cytotoxicity.
Cell Survival ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Hyaluronic Acid ; chemistry ; pharmacology ; Lipids ; Nanoparticles ; Particle Size ; Phosphatidylethanolamines ; chemistry ; pharmacology ; Retinal Pigment Epithelium ; drug effects ; Transfection

Ultrasonic thrombus ablation is a newly-developed technology for percutaneous arterial recanalization. An ultrasound angioplasty device is described here in detail. The device has an adjustable power output range and distal tip longitudinal displacement range. Experimental data suggest that this ultrasound device is significantly effective in ablating fresh thrombi.
Catheter Ablation ; instrumentation ; Equipment Design ; Expert Systems ; Thrombolytic Therapy ; Transducers ; Ultrasonography, Interventional ; Vibration

Objective: To observe the influence of Nie-pinching the spine manipulation on the excretory rate of urine D-xylose in the infants with diarrhea due to spleen deficiency, and to assess the clinical effects. Methods:Sixty infants in conformity with the diagnostic criteria of diarrhea due to spleen deficiency were randomly divided into a treatment group and a control group by the random digital table, 30 cases in each group. The infants in the treatment group were treated by Nie-pinching the spine manipulation and traditional infantile tuina, in addition to the routine basic treatment. The infants in the control group were treated by the same traditional infantile tuina in addition to the routine basic treatment. The infants in the two groups were treated once every day, 4 weeks as a course. Totally, the treatment was given for a course. The symptom integrals of spleen deficiency were used to assess the improvement in the symptoms. The colorimetry was used to determine the excretory rate of urine D-xylose. Results:In the comparison of the same group before and after the treatment, the differences in the global score of spleen deficiency symptoms and the excretory rate of urine D-xylose in the two groups were statistically significant (allP<0.01). After the treatment, the differences in the global score of spleen deficiency symptoms and the excretory rate of urine D-xylose between the two groups were all statistically significant (bothP<0.01). Conclusion:Chiropractics can reduce the integrals of spleen deficiency symptoms and elevate the excretory rate of urine D-xylose in the infants with diarrhea due to spleen deficiency, so as to enhance the therapeutic effects by alleviating the symptoms of spleen deficiency and the absorptive function of the small intestine.

Cinobufacino injection is purified from water extraction of the skin of Bufo bufo gargarizans, which has been widely used for various cancers in clinic with significant anti-tumor effects. Bufadienolides were regarded as the main active constituents of cinobufacino injection in previous reports. In present study, 6 bufadienolides were isolated and purified from Cinobufacino injection. Their structures were identified as 3-epi-ψ-bufarenogin (1), ψ-bufarenogin (2), 3-epi-arenobufagin (3), arenobufagin (4), 3-epi-gamabufotalin (5), and 3-oxo-arenobufagin (6), separately. Among them, 1 and 3 were new compounds, 5 and 6 were new natural products. Compounds 1, 2 and compounds 3, 4 were two pairs configuration isomers at C-3, separately.

Cinobufacino injection is a significant anti-tumor medicine for the treatment of various tumors in clinic, which was made from water extraction of the skin of Bufo bufo gargarizans. In present paper, HPLC-DAD-FT-ICR-MS method was used to identify the major bufadienolides in cinobufacino for the first time. Solid-phase extraction with dichloromethane and silica was used to enrich the total bufadienolides in cinobufacino. Based on the UV and high resolution MS/MS data, 33 bufadienolides were analyzed and characterized. Among them, eight compounds were identified by comparing with standard references unambiguously. This study elucidated the major bufadienolides in cinobufacino, which provided material foundation of cinobufacino and will be benefit for the further pharmacological research.