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Objective:To investigate the influencing factors of goiter in school-age children aged 8 to 10 in non-high iodine areas of Shijiazhuang City.Methods:In April 2018, 9 non-high iodine counties (cities) were selected as monitoring sites in Shijiazhuang City, and capacity proportional probability sampling (PPS) method was used. Each monitoring site was divided into five sampling areas according to five orientations: east, west, south, north, and middle, one township was selected from each area, one elementary school was selected from each township, and 40 school-age children aged 8 to 10 (balanced age, half males and half females) were selected from each school as respondents. Urine samples from any one time of children and drinking water samples from their village were collected, and urinary iodine and water iodine were detected by arsenic cerium catalytic spectrophotometry; the thyroid volume of children was measured by B ultrasound method; at the same time, the height and weight of children were measured and the body mass index was calculated. The influencing factors of goiter were analyzed by logistic regression analysis.Results:A total of 1 867 urine samples of school-age children were collected, and the median urinary iodine was 190.65 μg/L, which was in the suitable level of iodine. A total of 1 046 drinking water samples were collected, water iodine ranged from 0.11 to 87.91 μg/L, and the median water iodine was 3.01 μg/L. A total of 1 867 school-age children were tested thyroid, the median thyroid volume was 3.01 ml. The medians thyroid volume of boys and girls (928 and 939 cases) were 2.90 and 3.13 ml, respectively, the difference was statistically significant between sex ( U = 2.09, P < 0.05); the medians thyroid volume of children aged 8, 9, and 10 years old (622, 629, 616 cases) were 2.47, 2.87, and 3.13 ml, respectively, the differences were statistically significant among ages ( H = 203.96, P < 0.01); the medians thyroid volume of normal, overweight and obese children (1 231, 300, 336 cases) were 2.61, 3.05 and 3.16 ml, respectively, the differences were statistically significant among body mass index ( H = 65.55, P < 0.01). The results of multifactorial logistic regression analysis showed that female and obesity were risk factors of goiter in school-age children [odds ratio ( OR) = 2.08, 2.86, 95% confidence interval ( CI): 1.05 - 4.12, 1.39 - 5.88, P < 0.05]. Conclusion:Female and obesity are risk factors of goiter in school-age children aged 8 to 10 in non-high iodine areas of Shijiazhuang City.
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In the original publication, the label of Fig. 2C should be read as "GFAP/lectin/DAPI" not "DMP1/GFAP/lectin/DAPI".
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The blood-brain barrier (BBB) is a tight boundary formed between endothelial cells and astrocytes, which separates and protects brain from most pathogens as well as neural toxins in circulation. However, detailed molecular players involved in formation of BBB are not completely known. Dentin matrix protein 1 (DMP1)-proteoglycan (PG), which is known to be involved in mineralization of bones and dentin, is also expressed in soft tissues including brain with unknown functions. In the present study, we reported that DMP1-PG was expressed in brain astrocytes and enriched in BBB units. The only glycosylation site of DMP1 is serine89 (S89) in the N-terminal domain of the protein in mouse. Mutant mice with DMP1 point mutations changing S89 to glycine (S89G), which completely eradicated glycosylation of the protein, demonstrated severe BBB disruption. Another breed of DMP1 mutant mice, which lacked the C-terminal domain of DMP1, manifested normal BBB function. The polarity of S89G-DMP1 astrocytes was disrupted and cell-cell adhesion was loosened. Through a battery of analyses, we found that DMP1 glycosylation was critically required for astrocyte maturation both in vitro and in vivo. S89G-DMP1 mutant astrocytes failed to express aquaporin 4 and had reduced laminin and ZO1 expression, which resulted in disruption of BBB. Interestingly, overexpression of wild-type DMP1-PG in mouse brain driven by the nestin promoter elevated laminin and ZO1 expression beyond wild type levels and could effectively resisted intravenous mannitol-induced BBB reversible opening. Taken together, our study not only revealed a novel element, i.e., DMP1-PG, that regulated BBB formation, but also assigned a new function to DMP1-PG.
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Animals , Female , Male , Mice , Astrocytes , Cell Biology , Metabolism , Blood-Brain Barrier , Cell Biology , Metabolism , Cells, Cultured , Extracellular Matrix Proteins , Genetics , Metabolism , Glycosylation , Proteoglycans , Metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
In this paper,the recent advances in both biomedical sciences and higher medical education reform were reviewed and analyzed.Furthermore,we proposed and reconstructed the teaching system of biochemistry and molecular biology course in our university,including its teaching content,teaching methods,teacher team,teaching management,etc.The preliminary practice of this system has obtained significant positive effects on teaching quality and student performance.
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Tamoxifen and letrozole are two of the most effective pharmaceuticals mainly used in hormonal dependent) breast cancer therapy. A trace analytical method using ultra performance liquid chromatography/tandem mass spectrometry(UPLC-MS/MS) was developed to simultaneously detect these two drugs in both influent) and effluent of municipal sewage treatment plant. Sewage samples were passed through an Oasis HLB cartridge, then an amino solid phase extraction cartridge was connected with Oasis HLB cartridge. Target compounds) was firstly eluted with 6 mL methanol and then 3 mL methanol was used to elute the amino solid phase extraction cartridge, collected and pooled the elutants. After concentration, target compounds were separated) on a BEH C_(18)) column using a gradient elution profile with a mobile phase consisting of 0.1% formic acid aqueous) solution and acetonitrile and detected by an electro spray ionization tandem mass spectrometry with multiple reactions monitoring(MRM). Satisfactory linearity(R~2>0.997) was obtained over the range of 1.0-100 μg/L) and 0.1-100 μg/L for tamoxifen and letrozole, respectively, with limits of quantification(LOQ) of 1.0 and 0.1 ng/L. Mean recoveries of two target compounds(spiked in sewage samples at three concentration levels) ranged from 68.8% to 103.0%, with relative standard deviations(RSD) less than 15%. This method can be applied for the analysis of target drugs sewage samples.
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Objective:To study the effects of the conditioned medium (CM) from human lung adenocarcinoma cell line A549 on the viability and apoptosis of human umbilical vein endothelial cells (HUVECs) and the role of PI3K-Akt signaling pathway in the process. Methods:HUVECs were cultured with CM of A549 cells. Cell viability was detected by XTT assay. The morphological changes of HUVECs were analyzed by Hoechst 33258 staining and fluorescence microscopy. The apoptosis was detected by flow cytometry. Expression levels of total Akt and phosphorylated Akt were assessed by Western blotting. PI3K inhibitors wortmannin(WT)and Akt1 siRNA(siAkt1)were used to block PI3K-Akt signaling pathway. The mRNA transcription of Akt subtype was determined by RT-PCR.Results:A549 CM significantly increased cell viability after 24 h treatment (P=0.037) and inhibited apoptosis (P=0.001) of HUVECs. CM time-dependently activated phosphorylation of Akt. Akt was phosphorylated at 15 min after CM treatment and reached the peak at 30 min and then tended to decline. Both WT and siAkt1 blocked the effects of CM. Conclusion:The CM of A549 cells increased the survival and inhibit the apoptosis of HUVECs. Akt1 played a significant role in the process.
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Objective To investigate the expression and clinical significance of the estrogen receptor beta isoforms and survivin in thyroid tumors. Methods The pathological data of 125 patients with thyroid tumors were, collected from june 2003 to june 2007 in our institution, including thyroid carcinoma (86 cases), thyroid follicular adenoma (39 cases) and normal thyroid tissue (10cases). SP immunohistochemistry was used to measure the expressions of ERβ and survivin in the thyroid tumors. Results ERβ not only was detected in the thyroid epithelial cell plasma and nuclear, but also in the stroma. The positive rate of ERβ in cases with thyroid carcinoma, adenoma and normal thyroid tissue were 83.72 %, 51.28 % and 20.00 % respectively. There was significant difference between carcinoma and the adenoma or normal thyroid tissue (P<0.05). The positive rate of survivin in cases with carcinoma, adenoma and normal thyroid tissue were 59.30 %, 17.95 % and 0 respectively. There was significant difference between carcinoma and the adenoma or normal thyroid tissue (P<0.05). The expressions of ERβ and survivin correlated with TNM staging and lymph node metastasis of thyroid carcinoma (P<0.05). Conclusion ERβ may play a role in accelerating proliferation in the occurrence of the thyroid carcinoma. The expressions of ERβ and survivin are related to invasion and metastasis of thyroid carcinoma.
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The basic premise of syndrome essence discussion is the standardization of traditional Chinese medicine (TCM) syndrome type. However, there still exists confusion regarding the standardization of TCM syndrome differentiation treatment, and the guidelines for TCM syndrome differentiation could not really be used for guiding clinical treatment. This is mainly due to the inappropriate use of research ideas and methods. The fundamental research of TCM syndrome based on the differentiation and classification of diseases is the main method for studying the standardization of TCM syndrome type. The accuracy quantification of symptoms is the powerful guarantee for authenticity and reliability of the results from standardization study of syndrome type. The correct choice for statistical methods gives powerful technical support to determine the differentiation threshold. The unified scales, expert discussions and complex scientific theories are the best methods for current research on standardization of syndrome type. The correlation study of syndrome type and physicochemical indexes cannot reflect the syndrome type completely. It is supposed to establish the treatment principles according to the main pathological changes of diseases on the basis of the standardization of TCM syndrome differentiation.
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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: Troponin I (Tn I ) could inhibit the growth of vascular endothelial cells, inhibit neovascularization,through which to inhibit the development and metastasis of solid tumor. Similar to Tn I, TnC also exists in non-muscular tissue, but does it has the analogous activity of anticancer like Tn I ?OBJECTIVE: To explore the effect of recombinant human TnC (rhTnC) on the growth of human umbilical vein endothelial cells (HUV-EC) and mouse xenograft tumor.DESIGN: Controlled observation in vivo and in vitro.SETTING: Research Institute of Medicine, Chongqing K.E.W Pharmaceutical Co., Ltd. and Department of Biochemistry of Chongqing Medical University.MATERIALS: The 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. 100 Kunming mice either male or female of 15-22 g purchased from Chongqing Academy of Chinese Materia Medica. E.coli BL21 (DE3)pLysS/pET3b-TnC provided by Chongqing K.E.W Pharmaceutical Co., Ltd. HUV-EC (Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences).METHODS: Human TnC cDNA was obtained from human thymus cDNA library using PCR. The colony was cloned in E.coli and a bacterial strain of gene engineering E.coli BL21 (DE3) pLysS/pET3b-TnC was obtained, which could express hTnC. The recombinant human TnC (rhTnC) was purified with affinity chromatography of Ni-NTA agarose. ①In vitro cell experiment: HUV-ECs were seeded in the 96-well plates at density of 2×103 cells per well and co-cultured with rhTnC of 1, 5, 10, and 50 mg/L for 3 days. The absorption (A value) was detected with microplate reader at 540 nm and the inhibition rate of cell growth was calculated. Meanwhile, the 50% inhibiting dose (IC50 value) was assayed by LOGIT method. ②In vivo animal experiment: Ascites tumor (S-180) that had been inoculated for 7-8 days was harvested. The tumor cells were diluted to 1 ×1010 L-1 and 0.2 rnL was subaxillarily and intraperitoneally injected into each mouse (50 mice in each group). The next day, the mice were randomly divided into 5 groups: rhTnC 20 mg/kg group, rhTnC 10 mg/kg group, rhTnC 5 mg/kg group, Cyclophosphamide (Cy) group and control group with 10 mice in each group. The rhTnC 20,10 and 5 mg/kg groups were given administration at the corresponding doses, once a day for 7 days; 50 mg/kg Cy was given the Cy group one after an interval of day, and the same volume normal saline was given to the control group. One day after the last time of administration, all mice were killed and the tumor was harvested and weighed. The inhibition rate of tumor growth was calculated: tumor inhibition rate=[(Average weight of tumor in control group-Average weight of tumor in drug group)/Average weight of control group]×100%.MAIN OUTCOME MEASURES: Inhibition rate of rhTnC to HUV-EC proliferation in cell experiment in vitro and mouse xenograft tumor in animal experiment in vivo.RESULTS: ①In vitro cell culture showed that rhTnC suppressed HUV-EC proliferation in a dose-dependent manner (IC50=7.5 mg/L). ②Similar to the result of in vitro cell experiment, after intraperitoneal administration, the inhibition rate of rhTnC 5, 10, and 20 mg/kg groups was higher than that of control group (P ＜ 0.01); after subaxillary administration, the inhibition rate of rhTnC 5, 10, and 20 mg/kg groups was also higher than that of control group (P ＜ 0.05-0.01). There was no significant difference in the inhibition rate between two administration approaches (P ＞ 0.05).CONCLUSION: rhTnC is capable of inhibiting the proliferation of HUV-EC dose-dependently, and displays the activities of inhibiting the proliferation of HUV-EC and anti-tumor.
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Considering the character of biochemistry,this paper discussed teaching method and accounted for the importance of experiment teaching in biochemistry by some demonstrations including analyzing carefully for catching common ground;integrating theory with practice for helping understand and memorize;agitating the students' study interest etc.In addition,the importance of students was not to be ignored in teaching process.
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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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The key to deepen quality education in university is to integrate the education of humanities into speciality instruction.Research shows that the ways to combine them are varied: starting from knowledge,mental enlightenment, aesthetic pentration, interaction, classroom atmosphere creation, and anthropocentrism ideal promotion,etc.
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Objective To investigate the effects of inhibiting ubiquitin proteasome pathway(UPP) on proliferation of gastric carcinoma cells and the possible mechanism was discussed. Methods The gastric carcinoma cell strain SGC 7901 was treated with MG 132 to inhibit its UPP specially. The effect of growth suppression on cells was evaluated with MTT assay. Cell cycle and apoptosis were detected by flow cytometry(FCM). DNA fragment analysis was used for confirming the presence of apoptosis. The activity of telomerase was examined by TRAP PCR ELISA. Expression of p27kip1 was detected by immunocytochemical technique. Results MG 132 had great inhibitory effect on the growth of SGC 7901 cells. The FCM analysis showed that the ratio of G0/G1 phase of control group was (46.3?4.1)%, the ratio of G0/G1 phase of SGC 7901 cells treated with MG 132 increased to (72.1?5.0)% ( P
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Objective To investigate the expression and clinical significance of the estrogen receptor(ER) subsets ER?,ER? and Ki-67 in tissues of different thyroid diseases.Methods The pathological data of 93 patients with thyroid diseases were collected from Jun.2003 to Jun.2007,including thyroid carcinoma(56 cases),thyroid follicular adenoma(13 cases),nodular goiter(14 cases) and normal thyroid tissue(10 cases).S-P immunohistochemistry was performed to determine the expressions of ER?,ER? and Ki-67 in the thyroid tissues.Results ER? and ER? were expressed in the plasma and nuclei of thyroid epithelial cells,and ER? was also expressed in the stroma.The positive rates of ER? expression in cases with thyroid carcinoma,thyroid follicular adenoma,nodular goiter and normal thyroid tissue was 87.5%,69.2%,42.9% and 30.0%,respectively,with significant difference between carcinoma and goiter or normal thyroid tissue(P0.05).Conclusion ER? and ER? may play an aggravating effect on cell proliferation in the disease course of the thyroid carcinoma.The expressions of ER? and Ki-67 are related to invasion and metastasis of thyroid carcinoma.
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Objective:To develop a high cell-density fermentation process in micro DCU-400 fermentor of Pichia pastoris GS115 containing a recombinant vector pPICZ ?/endostatin.Methods:Cells were grown in the basal salts medium(BSM) at 30℃,pO 2≥30%,pH 5.3(growing) or pH 5.0(inducing),and the feed operation using 50% glycerol was performed when dissolved oxygen(pO 2) increased(after approximately 16-18h).It lasted 3-5hrs.When cell density(OD 600 ) reached 450,the inducement was began.After the transition from glycerol-methanol mixture to methanol,methanol from 0.03ml/min/L to 0.08ml/min/L in 8h was used to induce the endostatin expression.The inducement lasted for approximately 60h.Then centrifugation,the supernatant was collected.The expression was examined using 12% SDS-PAGE and ELISA.Results:In inducing phase,OD 600 reached 580(wet wt 275g/L).The production of endostatin is 27.2mg/L,exceeding that in spinner flask(16.3mg/L).Conclusion:The human endostatin had been successfully expressed in 15L fermentor.
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Objective:To explore the purification method of non- affinity chromatography for leptin expressed in Pichia Pastoris.Methods:Ion exchange chromatography (Sepharose Q fast flow) and hydrophobic interaction chromatography (Phenyl Sepharose 6 fast flow ) were used to purify human leptin in pH 7.5. Results: After purification by Q column,the purity of leptin increased from 42.3% to 89.6%.Subsequently, after hydrophobic interaction chromatography ,its purity reached 96.2%. In SDS-PAGE, leptin was shown as one specific band.Conclusion:The human recombinant leptin expressed by Pichia Pastoris yeast can be successfully purified by ion exchange chromatography plus hydrophobic interaction chromatography.