OBJECTIVETo establish a method for the determination of gossypol in cotton root bark.

METHODSamples were extracted with acetone, and determined on a C18 column with the mobile phase (Acetonitrile-0.2% phosphoric acid, 85:15) and UV-235 nm detector.

RESULTThe recovery rate was 97.6%, RSD 1.59% (n = 5).

CONCLUSIONThis method can be used for the determination of gossypol in cotton root bark and the content of gossypol in three different species has been determined.

Chromatography, High Pressure Liquid ; Gossypium ; chemistry ; Gossypol ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

Objective To investigate the effects of integrin β1 gene expression inhibited by short hairpin RNA (shRNA) on invasion of pancreatic carcinoma PANC1 cells in vitro,and investigate the mechanism.Methods The eukaryotic expression plasmid of shRNA targeting integrin β1 gene ( integrin β1 shRNA) and control eukaryotic expression plasmid shRNA (c-shRNA) was constructed and was transfected into PANC1 cells.The cells without plasmid transfection were used as control.The expressions of integrinβ1,MMP 2,MMP 9 mRNA and protein were detected by real-time PCR and Western blotting.The invasive ability of PANC1 cells was observed with Transwell cell culture chamber.Results Integrinβ1 mRNA expressions in integrinβ1 shRNA group,c-shRNA group and control group were 0.0029 ± 0.0004,0.0131 ± 0.0009,0.0138 ± 0.0005 ; the expressions of integrinβ1 protein were 0.0159 ± 0.0062,0.3215 ± 0.0126,0.3107 ±0.0094; the inhibitory rate of integrinβ1 mRNA and protein expression in integrinβ1 shRNA group was (78.6 ±7.2 ) ％ and (92.9 ± 3.2) ％ ( P ＜ 0.01 ).But there was no difference between the c-shRNA group and control group (P =0.2999).Number of penetrating cells in integrinβ1 shRNA group decreased from 52 ±5 to 21 ±4( P ＜ 0.01 ) ; the expression of MMP 2 and MMP 9 mRNA decreased from 0.592 ± 0.073,0.847 ± 0.069 to 0.102 ± 0.034,0.273 ± 0.071 ; the expression of M MP2 and MMP 9 protein decreased from 0.225 ± 0.046,0.416 ±0.081 to 0.059 ±0.013,0.106 ±0.022(P ＜0.05).Conclusions Recombinant integrinβ1 shRNA expression plasmid can effectively inhibit the expression of integrinβ1 gene and suppress the invasion of PANC1 cells in vitro by down-regulating MMP 2 and MMP 9 gene expression.

Objective To study the expression of p53,p16,PCNA protein in esophageal carcinoma and its relationship to sexual distinction,the location of disease,the biological level,the depth of invasion and lymph node metastasis.Methods 118 patients with esophageal carcinoma were included in the study,all of them were treated for the first time.p53,p16 and PCNA protein in the 118 cases of esophageal carcinoma were detected by immunohistochemical assay(SP technique). Results The positive expression of p53, p16, PCNA protein in 118 patients was 80 %(92/118),42%(50/118)and 97%(115/118),respectively.The positive expression of p53,PCNA protein were irrelated to the sexual distinction,the location of disease,the biological level,the depth of invasion and the lymph node metastasis.The loss of p16 was significantly related to the depth of invasion and the lymph node metastasis(P

Objective To investigate the protective effect of crocin on oxidative stress cell model of PC 12 cell and the effect of crocin on PI3K/Akt signal pathway,as well as further explore the mechanism of protective effect on model cells.Methods Cells were divided into control group,model group,crocin group and VE group.The cell survival rate was detected by MTT method,and the expression of mRNA and protein of PI3K/Akt were detected by RT-PCR and Western blot.Results With the crocin concentration in 0.625 μM and 5 μM,the cell survival rate increased in a dose-dependent manner.The average optical density rate of PI3K and Akt mRNA were 0.435±0.044 and 0.375 ± 0.034,and the PI3K and Akt protein were 0.378± 0.038 and 0.386± 0.043 of crocin group.Compared with the model group,the expression levels of PI3K/Akt increased in crocin group (P＜0.05).Conclusion These results indicate that the antioxidant and antiapoptosis effects of crocin are induced via increasing expression of PI3K and pAkt.

Objective To compare clinical features,pulmonary functions,chest imaging and prognosis between combined pulmonary fibrosis and emphysema syndrome (CPFE) and idiopathic pulmonary fibrosis(IPF) without emphysematous changes in elderly patients.Methods 88 elderly IPF patients in Beijing hospital from January 2000 to October 2012 were divided into CPFE (n=30) and IPF (n =58) groups according to the CT imaging.Clinical features,blood gas analysis,pulmonary function,chest CT and survival time were compared between the two groups.Results 30 CPFE patients with the mean age of(75.5 ±7.6) years and 58 IPF patients without emphysema with the mean age of(73.7±6.8) years were enrolled.The proportions of male patients,smoking history and mortality were higher in CPFE patients than in IPF group(86.7％ vs.63.8％,28 vs.36,76.7％ vs.43.1％,x2 =5.09,9.74,8.98,P＜0.05 or 0.01).CPFE patients had a higher force vital capacity(FVC) and total lung capacity(TLC) as compared with IPF group [(2.6±0.9) L vs.(2.1± 0.5) L,(5.4±1.9) L vs.(4.4±1.1) L,t=2.69,2.35,P＜0.01 or 0.05].There were no significant differences in forced expiratory volume in one second(FEV1) and the diffusion capacity for carbon monoxide(DLCO) between the two groups.The main type of emphysema by HRCT scan were centrilobular emphysema in CPFE patients.There were lower median survival time in CPFE group than in IPF group [(3.0±0.2) years vs.(4.0±1.0) years,x2=4.50,P＜0.05].Conclusions The majority of elderly CPFE patients are males and smokers.The lung volume is increased in elderly CPFE patients as compared with IPF patients.Centrilobular emphysema is the primary type in CPFE patients.The prognosis is worse in elderly CPFE patients than in IPF patients.

Objective To evaluate X-ray,CT and MRI findings of soft tissue infections in AIDS. Methods Three cases of soft tissue infections with AIDS were retrospectively analyzed by comparing the imaging findings with pathological results.All patients were performed MRI,X-ray was in 1 case,CT was in 1 case.Results Cellulitis was in 1 case:MRI showed extended thickening of subcutaneous tissues, ill-defined hypointense areas on T_1WI and hyperintensity on T_2WI,and reticular pattern on GRE. Necrotizing fasciitis was in 1 case:MRI showed obvious thickening of subcutaneous tissues and deep fasciae, abnormally increased signal intensity on T_1 and T_2WI.Fluid collections were within muscles and muscles interval on fat-suppressed T2 WI.Tuberculosis was in 1 case:CT demonstrated multiple low density areas in the subcutaneous tissues and clear peripheral rim enhancement.MRI appeared hypointense on T_1WI and hyperintensity on T_2WI,and peripheral rim enhancement following gadolinium injection.Conclusion Infections of soft tissue are common complication in patients with AIDS,radiology is important in early diagnosis and treatment planning in this population.

OBJECTIVETo investigate the expression of reversion-inducing cysteine-rich protein with Kazal motifs (RECK), matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor-C (VEGF-C) in colorectal cancer.

METHODSS-P immunohistochemistry was used to detect the expression of RECK, MMP-9 and VEGF-C in colorectal cancer specimen. Colorectal mucosal tissue at least 10 cm away from the tumor was used as control.

RESULTSThe positive expression of RECK in colorectal cancer was significantly lower than that in the controls(53.3% vs. 100%, P<0.05). Both MMP-9 and VEGF-C were over expressed compared with the controls(86.7% vs. 23.3% and 71.7% vs. 13.3% respectively, P<0.01). The expression of RECK was negatively correlated with that of MMP-9 and VEGF-C, and the expression of MMP-9 was positively correlated with that of VEGF-C. There were significant associations between the expression of these proteins and lymph node metastasis, distant metastasis, and TNM staging (all P<0.05).

CONCLUSIONSRECK expression is low in colorectal cancer, while MMP-9 and VEGF-C expressions are high. Combined testing of these 3 markers is important in the evaluation of tumor metastasis and invasion, and is helpful in the prediction of the prognosis.

Adult ; Aged ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Female ; GPI-Linked Proteins ; metabolism ; Humans ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; Prognosis ; Vascular Endothelial Growth Factor C ; metabolism

UNLABELLEDFrom large-scale sequence of human fetal liver cDNA library, we have obtained a full-length cDNA from an EST after further sequencing. It has been demonstrated by the alignment comparison with data base available that it is a novel member of Ubc family and got the number from GeneBank: UBF-F1 AF 294842.

AIM AND METHODSTo demonstrate its authenticity, UBF was amplified from the total RNA of human fetal liver and HL-60 cell line using RT-PCR, and the PCR products were further sequenced and compared with the original UBF sequence. To evaluate the expression level and subcellular location of UBF in human multiple tissues, in situ hybridization was carried out on the frozen section of human fetal multiple tissues and HL-60 cell line with DIG-labeled UBF cDNA probes.

RESULTSThe experimental results of RT-PCR and sequencing showed that the sequence of RT-PCR products were the same as the original UBF. The experimental results of in situ hybridization showed that UBF was expressed widely by human multiple fetal tissues and the expression level were very high in HL-60 cells.

CONCLUSIONIt is suggested that the special structure of UBF is authentic, and the expression profiling research of UBF shows that UBF is expressed widely by human multiple fetal tissues and the expression level is very high in HL-60 cells, implying that UBF plays the important function in the developing tissues and leukemia cells. It is also suggested that UBF may be functionally related with the nucleic-involving cellular activities based on the results of sub-cellular localizations.

Amino Acid Sequence ; Cloning, Molecular ; Gene Expression Profiling ; HL-60 Cells ; Humans ; Molecular Sequence Data ; Pol1 Transcription Initiation Complex Proteins ; genetics ; Reading Frames ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Ubiquitin-Conjugating Enzymes ; classification ; Ubiquitination

To investigate the function of exogenous unsaturated fatty acids in hyposmotic membrane stretch enhancement of muscarinic current (ICCh) in antral circular smooth muscle cells of guinea pig, we recorded the membrane current with the conventional whole cell patch-clamp technique. I(CCh) elicited by 50 micromol/L carbachol (CCh) at the holding potential of 20 mV under isosmotic condition was taken as control. Hyposmotic membrane stretch increased I(CCh) to 226.0+/-21.0%. When the cells were pretreated with 5 micromol/L arachidonic acid (AA), linoleic acid (LA) or oleic acid (OA), I(CCh)was inhibited to 3.8+/-0.6%, 35.2+/-0.8% and 66.6+/-0.6% respectively. Hyposmotic membrane stretch increased I(CCh) to 106.0+/-2.5%, 173.2+/-6.8% and 222.1+/-11.0% of the control respectively. Five micromol/L AA inhibited hyposmotic membrane stretch-enhanced I(CCh) by 51.2+/-3.8%, while the control I(CCh) under isosmotic condition was inhibited by 96.2+/-1.6%. The results suggest that unsaturated fatty acids inhibited I(CCh) and the inhibitory effect is more significant when the unsaturation degree is increased. However, the unsaturated fatty acids are not involved in the increase of I(CCh) induced by hyposmotic membrane stretch.
Animals ; Fatty Acids, Unsaturated ; pharmacology ; Guinea Pigs ; Membrane Potentials ; drug effects ; Myocytes, Smooth Muscle ; cytology ; physiology ; Osmotic Pressure ; Patch-Clamp Techniques ; Pyloric Antrum ; cytology ; physiology ; Receptors, Muscarinic ; physiology ; Sodium Chloride

Pseudoallergic reactions of Qingkailing injection (QKLI) was assessed by vascular hyperpermeability which were indicated by ear blue staining in ICR mice after single intravenous injection of QKLI mixed with Evans blue (EB) and skin blue spot formation in SD rats after intradermal injection of QKLI and intravenous injection of EB. In addition, QKLI-induced histamine, VEGF, TNF-alpha release was measured after ICR mice received the single dosing of QKLI iv. The mild vascular hyperpermeability characterized by ear blue staining could be observed in mice after intravenous injection of QKLI and EB. Intracutaneous injection of 50 micro L of test solution containing QKLI (25,50 microL) in rat back skin caused obvious local vascular hyperpermeability at the injection sites so as to result the larger diameters of blue spots than that in negative control group (P <0. 01). QKLI induced a significant increase of VEGF and a slight elevation of histamine in mice after intravenous administration, while TNF-alpha showed no change after QKLI iv. The results in this study indicated that both intravenous injection and intracutanous injection of QKLI could induce vascular hyperpemeability so as to cause pseudoallergic reaction in mice and rats. QKLI-induced pseudoallergic reaction may be associated with the release of histamine and VEGF.
Animals ; Drug Hypersensitivity ; blood ; etiology ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Histamine ; blood ; Injections ; Male ; Mice ; Rats ; Skin ; drug effects ; immunology ; Tumor Necrosis Factor-alpha ; blood ; Vascular Endothelial Growth Factor A ; blood