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Objective To explore the value of virtual touch tissue imaging quantification(VTIQ) and contrast-enhanced ultrasonography(CEUS) in the differential diagnosis of benign and malignant breast lesions with non-mass type. Methods A retrospective analysis of 85 cases by conventional ultrasound examination for the type of breast lesions in patients of image data,with pathologic results as the gold standard,analysis of US,VTIQ,CEUS and VTIQ joint CEUS for the breast type value in the differential diagnosis of benign and malignant lesions. Results There were 46 benign cases and 39 malignant cases in 85 cases of breast non-mass lesions. The internal shear wave velocity (SWV) and SWV ratio of breast non-mass benign lesions were significantly lower than that of malignant lesions(3. 43 ± 0. 59 m/s vs 4. 76 ± 0. 95 m/s,1. 73 ± 0. 43 vs 2. 67 ± 0. 77). The sensitivity,specificity and accuracy of VTIQ in the diagnosis of breast non-mass-like lesions were 76. 9％,80. 4％ and 78. 8％ respectively. Type of breast malignant lesions CEUS was high-enhanced enhancement compared with glandular tissue around the contrast agent into early enhancement and immediately wash-out and more for the nonuniformity,around the lesions or internal contrast enhancement of blood vessels,the sensitivity of CEUS in the diagnosis of mammary gland non tumor lesions, specific degrees and accuracy were 79. 5％,80. 4％ and 80. 0％ respectively. VTIQ type combined with CEUS in the diagnosis of breast mass lesions sensitivity and accuracy of the specific degree were 89. 7％,93. 5％ and 91. 8％ respectively,the joint diagnosis efficiency was significantly higher than single use of CEUS or VTIQ (P< 0. 05),CEUS and VTIQ diagnostic value were significantly higher than the US (P < 0. 05). Conclusion CEUS,VTIQ type in breast tumors in the differential diagnosis of benign and malignant lesions has good diagnostic value,VTIQ joint CEUS complementary advantages,more comprehensive diagnostic information available pathological changes,the type of breast disease diagnosis accuracy is improved.
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BACKGROUND: It demonstrated that combined application of thymosin α 1 (TM-α1) and interferon (IFN) can enhance the antiviral activity of IFN.OBJECTIVE: To obtain recombinant fusion protein of TM- α1 and consensus IFN α (IFN α -con) with double activity of antiviral activity and immunity enhancement.DESIGN, TIME AND SETTING: The in vitro contrast experiment was conducted in the Biochemical Laboratory of Research Institute of Medicine, Chongqing K.E.W Pharmaceutical Co., Ltd. from March 2003 to December 2004.MATERIALS: The fusion gene fragment (TM- α 1+IFN α -con) was synthesized by Shanghai Sangon Biological Engineering Technology And Service Co., Ltd, WISH cell, and vesicular stomatitis virus (VSV) was purchased from Institute of Biochemistry and Cell Biology, commercial products of IFN α 1b, IFN α 2a and TM- α 1 was served as reference substance.METHODS: The preference for E. coli of fusion sequence coding TM- α 1 and IFN α -con were cloned into expression vector of pET-22b(+) and expressed in BL21(DE3)-Codon plus-RP-X, which was purified by precipitation of (NH4)2SO4, hydrophobic interaction chromatography, anion-exchange chromatography, cation-exchange chromatography and molecular exclusion chromatography. The antiviral activity of fusion protein was tested by cytopathic-effect inhibition assay, and effect of fusion protein on lymphocyte proliferation was tested by cell proliferative assay.MAIN OUTCOME MEASURES: The specific activity of fusion protein and its biological activity in promoting lymphocyte proliferation.RESULTS: The fusion protein was expressed as a soluble form, accounting for over 20% of the total cell protein in E. coli, which approached 96% after purification. The antiviral activity of fusion protein was superior to IFN α 1b and IFN α 2a. However, the activity of fusion protein for promoting lymphocyte proliferation was similar to TM- α 1.CONCLUSION: The fusion protein of TM- α 1 and IFN α -con expressed in E. coli has both effects on anti-virus and promoting lymphocyte proliferation.
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This study aimed to obtain recombinant fusion protein of thymosin alphal(TM-alpha1) and consensus IFNalpha (IFNalpha-con) which have bath TM-alpha1 and IFNalpha-con activities. The DNA sequence for the fusion protein was cloned into expression vector of pET-22b (+) and expressed in BL21 (DE3)-Codon plus-RP-X. The expressed product (TM-alpha1-IFN-con) was soluble, and amounted to more than 20% in total proteins of E. coli. By precipitation of (NH4)2SO4, hydrophobic interaction chromatography (HIC, Phenyl Sepharose 6 Fast Flow), anion-exchange chromatography (Q Sepharose Fast Flow), cation-exchange chromatography (SP Sepharose Fast Flow) and gel filtration (Sephadex G-75), it was purified to more than 96% purity. The activity of fusion protein for antivirus was tested by cytopathic-effect inhibition assay and activity for promoting lymphocyte proliferation was tested by cell proliferative assay. The activity for antivirus was higher than commercial IFNalpha1b and IFNalpha2a and activity for promoting lymphocyte proliferation was similar to commercial TM-alpha1. The fusion protein had better effect for anti-HBV in vitro, its effect was stronger than combination of IFNalpha and TM-alpha1 and cell toxicity was less than combination of IFNalpha and TM-alpha1. The above results show that it has effect bath antivirus of IFNalpha and promoting lymphocyte proliferation of the soluble fusion protein expressed in E. coli.
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Animals , Humans , Mice , Amino Acid Sequence , Antiviral Agents , Pharmacology , Base Sequence , Escherichia coli , Genetics , Metabolism , Interferon Type I , Genetics , Interferon-alpha , Molecular Sequence Data , Recombinant Fusion Proteins , Genetics , Pharmacology , Recombinant Proteins , Thymosin , GeneticsABSTRACT
BACKGROUND:As a by-product in sugar industry, sugar cane wax has been widely used in non-medical field. Some researches indicate that sugar cane wax plays a great role in reducing blood cholesterol; however, the therapeutical effect and clinical application should be studied further.OBJECTIVE: To separate the high-class eicosanoic acid and the high-class alkanols, which are suitable for medical application, and further to observe the effect of them on reducing blood cholesterol of model rets with hyperlipemia.DESIGN: Randomized control animal study.SETTING: Pharmacological Institute, Chongqing Kangerwei Pharmaceutical Co., Ltd.MATERIALS.: The experiment was carried out in the Pharmacological Institute of Chongqing K.E.W Pharmaceutical Go.,Ltd. From April 2005 to January 2006. A total of 65 adult female Wistar rats, aged at 3-6 months, weighing 180-220 g, of SPF grade, were provided by Experimental Animal Center of Chongqing Institute of Traditional Chinese Medicine. Raw sugar cane wax was provided by Beijing Jiade Hongsheng Biochemical Technology Co., Ltd. High-class alkanols C26,C28,C30, C32 and high-class eicosanoic acid C28, C30, C32, C34 were provided by Sigma Company (standard materials of gas phase chromatography), and other reagents were national analytical pure.METHODS: ① Sugar cane wax was extracted from raw sugar cane wax with ethanol and other organic solution and separated from the mixture of high-class eicosanoic acid and the mixture of high-class alkanols with saponification and calcification. Main components were analyzed with gas phase chromatography. The main components of high-class alkanols were C26, C28, C30 and C32 and the main components of high-class eicosanoic acid were C28, C30, C32 and C34, ② Based on references, rats were fed in 3 days and randomly divided into blank group (n =10) and experimental group (n =55).And then, all rats were cut off their tails to collect blood and the triacylglyoerol (TG), total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) were measured with automatic biochemistry analyzer. Rats were fed with common granule feeds in blank group or with high-lipid feeds (containing 0.1 mass fraction of oiliness, 0.1 mass fraction of yolk powder, 0.01 mass fraction of cholesterol, 0.002 mass fraction of pig's gall salt, 0.788 mass fraction of common feeds) in experimental group. All rats ate and drank freely. Seven days later, blood was collected again from tail tip to measure the contents of TG, TC and HDL-C. Based on level of serum TG, rats in the experimental group were randomly divided into 5subgroups (n =11): negative control group, low-dosage high-class alkanols group, high-dosage high-class alkanols group,Iowdosage high-class eicosanoic acid and high-dosage high-class eicosanoic acid group. Rats in low-dosage and high-dosage high-class alkanols groups were perfused with 5 and 50 mg/(kg·d) high-class alkanols; meanwhile, rats in low-dosage and high-dosage high-class eicosanoic acid groups were perfused with 20 and 200 mg/ (kg·d) high-class eicosanoic acid. Rats in negative control group and blank group were perfused with the same volume of 0.3% carboxymethylcellulose sodium and distilled water, respectively, once a day for successive 30 days. At 16 hours after the last administration, rats were anesthetized to collect blood from heart to measure contents of TG, TC and HDL-C in serum.MAIN OUTCOME MEASURES: ① Percentage of main component in separated mixtures of high-class eicosanoic acid high-class alkanols; ② levels of serum cholesterol, HDL and TG.RESULTS: A total of 65 experimental rats were involved in the final analysis. ① Gas phase chromatography suggested that the content of C28 high-class alkanols was the most (73.6%), and other three kinds of high-class alkanols were counted for 5.3% (C26), 6.2% (C30) and 5.1% (C32), respectively. The total quantity was 90.2%. In the mixture of high-class eicosanoic acid, content of C28 high-class eicosanoic acid was the most (46.6%) and the other three kinds of high-class eicosanoic acid were counted for 16.7% (C30), 6.8% (C32) and 9.3% (C34), respectively. The total quantity was 79.3%. ②Levels of serum TC were (1.46±0.27), (1.66±0.33), (1.44±0.25) and (2.16±0.52) mmol/L in high-dosage and Iow-dosage high-class alkanols groups and high-dosage and Iow-dosage high-class eicosanoic acid groups, respectively, which were lower than those in negative control group [(2.52±0.83) mmol/L, P＜0.01]. Levels of HDL-C were (0.73±0.09), (0.71±0.07), (0.79±0.10) and (0.70±0.08) mmol/L in the four treatment groups, respectively, which were higher than those in negative control group [(0.58±0.13) mmol/L, P＜0.05-0.01].CONCLUSION: The high-class alkanols and the high-class eicosanoic acids separated from sugar cane wax made in China significantly have the activity of reducing blood cholesterol; however, the effect on decreasing TG is not obvious.
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BACKGROUND: Leptin is a hormone produced predominantly by adipocytes and has a variety of physiological functions. It has been a hot spot in energy metabolism research. However, leptin presently used is usually produced by E coli, in which leptin cDNA is expressed and is in the form of insoluble inclusion body. Therefore, extremely complicated reactivating procedure is needed to obtain biologically activated human leptin.OBJECTIVE: To explore the condition for purification of leptin in non-affinity chromatography in order to obtain soluble human leptin.DESIGN: An observational experiment.SETTING: Molecular laboratory of biochemical department in a medical college.MATERIALS: The strong anion exchanger sepharose Q and hydrophobic phenyl sepharose 6 were used in different conditions for removal of as many contaminants as possible.METHODS: The supernatant of pichia pastoris yeast culture solution was first purified through Q column chromatography, the protein was collected and was further purified through hydrophobic support with 1 mol/ L (NH4) 2SO4.MAIN OUTCOME MEASURES: Gel scanning revealed that the purity of human leptin was 42. 3% before purification, 89.6% after Q column chromatography and 96.2% after hydrophobic interaction chromatography.RESULTS: The post-purification product presented a s. ingle band in SDS-PAGE. Gel scanning revealed that the purity of human leptin was 42.3% before purification, 89.6% after Q column chromatography and 96.2% after hydrophobic interaction chromatography.CONCLUSION: The combined use of strong anion exchange chromatography and hydrophobic interaction chromatography can effectively purify leptin expressed by pichia pastoris yeast and the purity is identical to that of nickel affinity column chromatography. It provides reliable evidence and method for possible manufacture of human leptin and lays experimental basis for leptin-related research.
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Objective:To investigate the radiosensitivity enhancement effect of COX-2 inhibitor-Nimesulide on breast cancer cell line(MCF-7) in vitro and to initially disclosure its mechanism.Methods:The clonogenic assay was performed to determine the radiosensitizing effect of Nimesulide on MCF-7 cell line.The morphological changes of MCF-7 were observed by electron microscopy.The apoptosis and expressions of Bcl-2 and Bax were measured by flow cytometry(FCM).Results: Nimesulide at different concentration significantly enhanced the radiosensitivity of MCF-7 cells.in a dose-dependent manner,under the combined treatment of Nimesulide and radial the protein expression of Bax was up-regulated,while that of Bcl-2 was inhibited and the apoptpsis increased significantly.Conclusion:Cox-2 inhibitor could obviously enhance the radiosentivity of human breast cancer cell line MCF-7.
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Objective:To study the effect of Hypoxia-inducible factor-1 alpha(HIF-1?)on the radiosensitivity of rat cervical carcinoma and discuss the possible mechanism.Methods:The transplantation tumor model of cervical carcinoma was established. 77 rats were randomly divided into five groups:control group,the large fraction irradiation group,the small fraction irradiation group,the large fraction irradiation combining HBO group and the low dose irradiation combining HBO group.Expressions of HIF-1? and bcl-2 in transplantation tumor after treatment were detected by immunohistochemical staining,and the tumor inhibitory rate by the change of tumor volume.The correlation between expressions of HIF-1 ? and bcl-2(and tumor inhibitory rate)was also observed.Results:Expression of HIF-1 ? and carcinoma growth inhibitory rate showed a reverse correlation(r=-5.147,P
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Objective:To investigate the inhibitory effect of cyclooxygenase inhibitor acetylsalicylic acid(ASA),and a selective COX-2 inhibitor(Nimesulide) on the growth of human breast cancer cell line MDA-MB-231 in vitro.Methods:The inhibitory effect was detected by MTT assay.Immunohistochemical assay was performed to examine the expressions of COX-2 and c-erbB-2 in SKOV3 cell line treated by NSAIDs.Results:A decrease in cell number compared with controls was observed in all of the cell line treated with ASA and Nimesulide.It was a dose-dependent inhibition of cell proliferation.COX-2 was expressed positive in MDA-MB-231 and the expression of cerbB-2 was found to decrease by immunohistochemical assay.Conclusion:NSAIDs can inhibit the growth of human breast cancer cell line MDA-MB-231 in vitro.