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1.
Article in Chinese | WPRIM | ID: wpr-1015604

ABSTRACT

Betulinic acid (BA) exerts protective effects on organs in septic animals. However, whether BA can improve cardiac function in sepsis and the underlying mechanism remain unclear. Here, male Sprague-Dawley rats were pretreated with BA (25 mg/ kg/ d, i. g.) for 5 days and then intraperitoneally injected with lipopolysaccharide (LPS, 10 mg/ kg). The rats were anesthetized to determine transthoracic echocardiography using a high-resolution imaging system for small animals after they were treated with LPS for 6 h. Histopathologic alterations were examined by HE staining. Myocardial injury markers (cTnI and CK-MB) and inflammatory factors (TNF-α, IL-1β and IL-6) in the serum were measured by the enzyme-linked immunosorbent assay. Autophagy-related proteins (p62 and LC3 Ⅱ) and AKT-modulated autophagy pathways in the myocardium were determined by Western blotting. Pretreatment with BA markedly improved left ventricular ejection fraction (EF) and fraction shortening (FS) (P<0. 05), improved myocardial histomorphology, and significantly inhibited cTnI, CK-MB, TNF-α, IL-1β and IL-6 (P<0. 05) in the septic rat serum. BA markedly decreased p62 (P<0. 01), increased LC3 Ⅱ (P< 0. 001), and significantly down-regulated p-AKT (Thr308), p-AMPKα (Ser485/ 491), p-mTOR (Ser2448) and p-S6K (Thr389) (P<0. 05), while markedly up-regulated p-AMPKα (Thr172) and pULK1 (Ser317) (P<0. 01) in septic rat hearts. The findings indicate that BA can attenuate sepsis-induced myocardial dysfunctions associated with down-regulating autophagy inhibiting pathways mediated by AKT/ mTOR and AKT/ AMPK pathways.

2.
Article in Chinese | WPRIM | ID: wpr-973407

ABSTRACT

Objective To investigate the effect of gaseous effluent from the six generator sets on the radiation level of the surrounding terrestrial environment in Daya Bay Nuclear Power Base after the operation of Ling’ao Nuclear Power plant. Methods The radiation level in the peripheral environment of the Base was monitored using the thermoluminescence dosimeter (TLD). Twenty-five monitoring sites were set around the Base to investigate the variation of radiation level over a long period of time by collecting the TLDs every three months. Results From 2011 to 2020, the annual γ dose rate of the 25 sites ranged from 76.7 to 207.1 nGy/h, with an average value of (123.3 ± 5.7) nGy/h and a relative deviation of 2%-12%. The TLD monitoring and instantaneous measuring results of γ dose rate were consistent with the survey of the State Environmental Protection Administration in the 20th century and the baseline level prior to the operation of the nuclear power plant. Conclusion There are great differences in natural environmental radiation level across the TLD monitoring sites. The overall environmental γ radiation level within 50 km of the nuclear power base remains unchanged. The emission of gaseous effluent from the operation of the nuclear power plant does not have a cumulative impact on the radiation level of surrounding environment.

3.
Chinese Journal of Radiology ; (12): 288-292, 2021.
Article in Chinese | WPRIM | ID: wpr-884425

ABSTRACT

Objective:To evaluate the safety and effectiveness of the accurate puncture during sacral neuromodulation (SNM) guided with 3D printing navigation template based on reconstruction techniques using fusing sacral CT and MRI images.Methods:Totally 42 patients operated with SNM were selected in Renji Hospital, School of Medicine, Shanghai Jiaotong University from July 2016 to August 2017. The patients were randomly divided into control group ( n=22) and experimental group ( n=20) using random number table. The conventional cross-positioning technique under X-ray was used for puncture during SNM in the control group. While in the experimental group, the sacral CT and MRI images were fused for reconstruction and design of the navigation template, printed by 3D technique for the puncture in SNM. The times of punctures, the average time for puncture operation, the time of intraoperative testing of the stimulator device, the minimum onset voltage of the stimulator, the X-ray radiation dose, postoperative curative effect (rate of secondary transformation) and the incidence rate of complications were compared between the two methods using independent-simple t test or χ 2 test. Results:Compared to control group, fewer times of punctures, shorter time needed for puncture operation, shorter time of intraoperative testing of the stimulator, smaller radiation dose and minimum effective voltage were found in the experimental group ( P<0.05). There were 15 and 16 patients who completed the secondary transformation in the control group and experimental group, and there was no significant difference between the two groups (χ2=0.757, P=0.384). There were 3 cases of complications in the control group, including 2 cases of infection and 1 case of bleeding, while no complications in the experimental group. Conclusions:CT and MRI images fusion reconstruction-guided 3D printing navigation template can help perform accurate and safe punctures in SNM. Compared to conventional puncture positioned under X-ray, it can effectively improve the puncture efficiency, and reduce the radiation dose in the operation.

4.
Article in 0 | WPRIM | ID: wpr-832070

ABSTRACT

Objective@#We conducted a meta-analysis of randomized controlled trials to explore whether vitamin D supplementation is beneficial for symptom improvement in children with autism spectrum disorder. @*Methods@#We systematically searched the PubMed database, EMBASE, Cochrane Library, Web of Science, Sino-Med, Wanfang Data, and China National Knowledge Infrastructure mainly up to September 2019. Using a fixed effects model, we calculated the standard mean difference with 95% confidence interval. Furthermore, we analyzed baseline serum 25-hydroxyvitamin D levels and outcome scores including the Social Responsiveness Scale and Child Autism Rating Scale scores after vitamin D supplementation. @*Results@#There was no significant difference in baseline serum 25-hydroxyvitamin D levels among 203 children included from three studies in the meta-analysis. After vitamin D supplementation, the outcome scores in the experimental group were dramatically elevated compared with those in the control group (p = 0.03). @*Conclusion@#Vitamin D supplementation improves the typical symptoms of autism spectrum disorder, as indicated by reduced Social Responsiveness Scale and Child Autism Rating Scale scores; thus, it is beneficial for children with autism spectrum disorder.

5.
Article in Chinese | WPRIM | ID: wpr-864158

ABSTRACT

Objective:To analyze the serum 25-hydroxyvitamin [25(OH)D] nutritional status of children with febrile seizures in Luzhou area of Sichuan Province, and the relationship of 25(OH)D with gender, age and the local season, so as to provide reference for vitamin D supplementation and prevention of febrile seizures in children in this area.Methods:One hundred and sixty-seven children diagnosed with febrile seizures in the Affiliated Hospital of Southwest Medical University from January 2015 to July 2018 were enrolled in the febrile seizures group, and 170 children aged 0-8 years who underwent health examinations in the outpatient department in the same period were included in the healthy control group.The serum total calcium, serum 25(OH)D and hemoglobin level of children with febrile seizures were analyzed.The correlation of febrile seizures with the level of serum 25 (OH)D was evaluated from the aspects of children gender, age and season.Results:(1) The serum 25(OH)D level of healthy children [(40.6±3.07) μg/L] was significantly higher than that of the children with febrile seizures [(27.18±6.68) μg/L], and the diffe-rence was statistically significant ( t=3.15, P=0.03). The serum 25(OH)D level in children with febrile seizures decreased with age.There was no deficiency found in serum total calcium and hemoglobins of all children with febrile seizures.(2) The incidence rate of febrile seizures was the highest in January (35 cases, 20.96%) and in 2-year-old toddlers (72 cases, 44.91%). Boys were significantly more susceptible to febrile seizures than girls (1.73∶1.00). (3) The serum 25(OH)D level of children with febrile seizures in the region decreased in winter and summer (27.47-30.37 μg/L), and increased in spring and autumn (31.58-35.13 μg/L). The serum 25(OH)D level of children in winter [(27.47±1.80) μg/L] was statistically significantly different from that in spring [(31.58±1.31) μg/L] and in autumn [(35.13±3.93) μg/L] (all P<0.05). The serum 25(OH)D level in children with febrile seizures were inversely proportional to the high temperature in summer [T>35 ℃, 25(OH)D<30 μg/L]. The 25(OH)D level showed a downward trend when the temperature exceeded 35 degrees Celsius.The optimum temperature for children in this area to absorb vitamin D by sun-irradiation was 25-35 ℃. Conclusions:The occurrence of febrile seizures in children of all ages in Luzhou is closely related to the deficiency of vitamin D. Two-year-old boys are prone to be attacked by febrile seizures in January of each year.In addition to winter, children, especially those over 2 years old, are advised to supply vitamin D in high temperature periods in summer when outdoor activities are reduced.

6.
Article in English | WPRIM | ID: wpr-1010506

ABSTRACT

Genome sequencing projects revealed massive cryptic gene clusters encoding the undiscovered secondary metabolites in Streptomyces. To investigate the metabolic products of silent gene clusters in Streptomyces chattanoogensis L10 (CGMCC 2644), we used site-directed mutagenesis to generate ten mutants with point mutations in the highly conserved region of rpsL (encoding the ribosomal protein S12) or rpoB (encoding the RNA polymerase β-subunit). Among them, L10/RpoB (H437Y) accumulated a dark pigment on a yeast extract-malt extract-glucose (YMG) plate. This was absent in the wild type. After further investigation, a novel angucycline antibiotic named anthrachamycin was isolated and determined using nuclear magnetic resonance (NMR) spectroscopic techniques. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis and electrophoretic mobility shift assay (EMSA) were performed to investigate the mechanism underlying the activation effect on the anthrachamycin biosynthetic gene cluster. This work indicated that the rpoB-specific missense H437Y mutation had activated anthrachamycin biosynthesis in S. chattanoogensis L10. This may be helpful in the investigation of the pleiotropic regulation system in Streptomyces.


Subject(s)
Anti-Bacterial Agents/pharmacology , Antioxidants/pharmacology , Bacterial Proteins/genetics , Multigene Family , Mutagenesis, Site-Directed , Streptomyces/metabolism
7.
Article in Chinese | WPRIM | ID: wpr-700767

ABSTRACT

Objective A variety of secondary metabolites can be obtained after the fermentation of medicinal fungi.The con-tent of these metabolites is much higher than that of cultivated medicinal fungi. We aimed to preliminarily study on the apoptosis of fer-mented Inonotus obliquus(FIO) on hepatocellular carcinoma HepG-2 cells. Methods The experiment was divided into 3 groups:the control group(The same volume of drug-free medium),the UIO group(200 mg/L) and the FIO group(200 mg/L). The contents of polysaccharides,terpenoids and polyphenols in unfermented Inonotus Obliquus (UIO) and FIO were determined by sulfuric acid phenol method,Folin-Ciocalteu method and colorimetric method. Effects of Fermented Inonotus obliquus on apoptosis of HepG-2 Cells were measured by MTT Method and Flow Cytometry. Results The contents of polysaccharides,terpenoids and polyphenols were sig-nificantly increased in FIO group [(10.140±0.849)mg/mL,(1.774±0.001)mg/mL, (9.979±0.022)mg/mL] compared with thatin UIO group [(7.161±0.305)mg/mL,(1.358±0.004)mg/mL,(6.314±0.237)mg/mL](P<0.01). Both UIO and FIO group induced apoptosis of HepG-2 cells according to Annexin V/PI double staining. Compared with the control group(4.3%),the apoptosis rate in-creased in UIO and FIO group (23.14%,27.37%) (P<0.05). In addition, the apoptosis rate was higher in FIO group than that in UIO group (P<0.05). Compared with the control group, the ratio of G0/G1phase cells was significantly increased and the ratio of S phase and G2/M phase was significantly decreased in UIO and FIO group (P<0.01). G0/G1phase cell ratio differences were statistically signif-icant compared with the UIO group (P<0.05) Conclusion FIO can better induce the apoptosis of HepG-2 cells than UIO.

8.
Tropical Biomedicine ; : 570-576, 2016.
Article in English | WPRIM | ID: wpr-630842

ABSTRACT

This study was designed to assess the immune protective effects of the vaccine strain of a precocious line of Eimeria necatrix with different doses and at different immunization times. The immunizations had a negative effect on weight gains of chickens to a certain degree but could be compensated during the “compensatory growth period” after immunity was established in the chickens. The number of oocysts excreted was positively correlated with the immunization dose. All the immunized chickens, whether they were immunized once or twice or immunized with different doses of sporulated oocysts, were able to resist attack from 1x105 virulent sporulated oocysts of E. necatrix. The lesion scoring showed that no significant difference existed in the chicken groups immunized with different doses (300 and 600) of sporulated oocysts. However, a difference existed in the immune homogeneity established in the different immunized groups, and two artificial immunizations were superior to one artificial immunization, indicating that two could extend the duration of oocyst excretion and allow more chances for the immunized chickens to become repeatedly infected.

9.
Clinics ; Clinics;68(1): 75-80, Jan. 2013. graf, tab
Article in English | LILACS | ID: lil-665921

ABSTRACT

OBJECTIVE: Recent studies have shown that circulating microRNAs might be useful, novel biomarkers for the diagnosis of acute myocardial infarction. The aims of this study were to evaluate the expression of cardiac-specific miRNAs (miR-1, -133a, -208b, and -499) in patients with acute myocardial infarction and to compare the diagnostic values of these miRNAs with that of cardiac troponin T. METHODS: Sixty-seven plasma samples obtained from patients with acute myocardial infarction and 32 plasma specimens collected from healthy volunteers were analyzed in this study. The levels of cardiac-specific miRNAs (miR-1, -133a, -208b, and -499) were measured by quantitative reverse transcription-polymerase chain reaction, and the concentrations of plasma cardiac troponin T were measured using electrochemiluminescence-based methods and an Elecsys 2010 Immunoassay Analyzer. RESULTS: The levels of plasma miR-1, -133a, -208b, and -499 were significantly higher in acute myocardial infarction patients (all p<0.001) than in healthy volunteers. The expression of the cardiac-specific miRNAs in acute myocardial infarction patients decreased to close to the baseline levels at the time of hospital discharge (all p>0.05). There were no correlations between the levels of the four circulating miRNAs and the clinical characteristics of the study population (all p>0.05). Furthermore, receiver operating characteristic curve analyses showed that the four plasma miRNAs were not superior to cardiac troponin T for the diagnosis of acute myocardial infarction (all p>0.05). CONCLUSION: Our results demonstrate that circulating miR-1, -133a, -208b, and -499 may be useful biomarkers in acute myocardial infarction patients but that these miRNAs are not superior to cardiac troponin T for the diagnosis of acute myocardial infarction.


Subject(s)
Aged , Female , Humans , Male , Middle Aged , MicroRNAs/blood , Myocardial Infarction/diagnosis , Troponin T/blood , Biomarkers/blood , Epidemiologic Methods , Immunoassay , Luminescent Measurements , Myocardial Infarction/genetics , Predictive Value of Tests , Reference Values , Reverse Transcriptase Polymerase Chain Reaction
10.
Article in Chinese | WPRIM | ID: wpr-302179

ABSTRACT

The objective of this study was to observe the expression of hoxc4 and hoxc6 genes in the process of differentiation of hematopoietic stem cell (HSC) to colony forming unit-T Lymphocyte (CFU-TL) in vitro. and to explore the possible mechanism of HCMV-induced maldevelopment of human cord blood CFU-TL on genetic level through effecting the differentiation progress by human cytomegalovirus (HCMV) with and/or all-trans retinoic acid (ATRA), Normal CFU-TL culture was used as blank control. After detection with MTT, mRNA expression levels in the human cord blood CFU-TL hoxc4 and hoxc6 genes following HCMV infection and ATRA treatment were detected by fluorogenic quantitative reserve transcription polymerize chain reaction (FQ-RT-PCR) method. HCMV of 10(6) plaque formation unit (PFU)/ml was diluted to 0.1 ml 10(5) PFU/ml and added into the infected group. The results showed that the expression of hoxc4 and hoxc6 genes in the differentiation process increased slightly on day 3, and were up to the most on day 7 (p < 0.05), while became lower on day 12 respectively in normal group, HCMV group and ATRA group. Compared with the expression of hoxc6, the expression of hoxc4 was obviously higher in each group (p < 0.05). Compared with the expression of hoxc4 and hoxc6 genes in normal group, the expressions of hoxc4 and hoxc6 in ATRA group were up-regulated remarkably (p < 0.05), while the expressions of hoxc4 and hoxc6 in group HCMV were down-regulated (p < 0.05). It is concluded that the regular expression of hoxc4 and hoxc6 genes mRNA appeared in each group. A positive co-relationship exits between hoxc4/hoxc6 genes and lymphocytic progenitor hematopoiesis. Compared with the expression of hoxc6 gene, the expression of hoxc4 gene is obviously higher in each group. HCMV can down-regulate the expression of hoxc4 and hoxc6 genes and lead to suppression effect on cell morphology, which confirms that the normal hematopoietic lineage determination and maturation rely on the stable and consistent expression of homeobox gene. At the same condition, ATRA (6 x 10(-8) mol/L at 60 nmol/ml) can up-regulate hoxc4 and hoxc6 genes expression. ATRA can up-regulate the expression of hoxc4 and hoxc6 genes.


Subject(s)
Humans , Cell Line , Cell Proliferation , Cytomegalovirus , Genetics , Cytomegalovirus Infections , Genetics , Homeodomain Proteins , Genetics , Lymphoid Progenitor Cells , Cell Biology , Tretinoin , Pharmacology
11.
Chinese Journal of Oncology ; (12): 254-257, 2009.
Article in Chinese | WPRIM | ID: wpr-293138

ABSTRACT

<p><b>OBJECTIVE</b>To observe the influence of endostar alone or in combination with cisplatin on tumor growth and metastasis, as well as the inhibition of angiogenesis and lymphangiogenesis in nude mouse models of human cervical cancer.</p><p><b>METHODS</b>HeLa cells were inoculated subcutaneously into the hind flank region of female nu/mice to establish xenograft models. The nude mice were randomly divided into 5 groups: (1) sodium chloride (as control); (2) cisplatin alone; (3) endostar alone; (4) cisplatin plus endostar (10 mg/kg); (5) cisplatin plus endostar (20 mg/kg). The course of all the treatments lasted for 4 weeks. The tumor growth and lymph node metastasis were observed. Immunohistochemical staining was employed to detect the angiogenesis and lymphangiogenesis.</p><p><b>RESULTS</b>(1) Either endostar alone or endostar with cisplatin inhibited the tumor growth significantly than cisplatin and NS (P < 0.05). (2) The rates of lymph node metastasis in the endostar (20 mg/kg) with cisplatin, the endostar (10 mg/kg) with cisplatin, the endostar, the cisplatin and the NS groups were 0 (0/8), 12.5% (1/8), 12.5% (1/8), 62.5% (5/8) and 75.0% (6/8) (P = 0.002), respectively. (3) The MVD of tumor tissue in these five groups were 10.88 +/- 1.38, 10.25 +/- 1.22, 10.83 +/- 2.29, 15.58 +/- 2.31 and 22.08 +/- 1.93, respectively (P < 0.05). The MLD were 5.00 +/- 0.63, 5.17 +/- 0.75, 6.00 +/- 0.63, 14.33 +/- 1.63 and 13.67 +/- 1.21, respectively (P < 0.05).</p><p><b>CONCLUSION</b>Endostar can reduce the tumor growth and metastasis by inhibiting angiogenesis and lymphangiogenesis in nude mouse model of human cervical cancer.</p>


Subject(s)
Animals , Female , Humans , Mice , Angiogenesis Inhibitors , Pharmacology , Antineoplastic Agents , Pharmacology , Cisplatin , Pharmacology , Endostatins , Pharmacology , HeLa Cells , Lymphangiogenesis , Lymphatic Metastasis , Lymphatic Vessels , Mice, Inbred BALB C , Mice, Nude , Microvessels , Neoplasm Transplantation , Neovascularization, Pathologic , Random Allocation , Tumor Burden
12.
Article in Chinese | WPRIM | ID: wpr-329284

ABSTRACT

The IVD (In Vitro Diagnostic)Medical Devices are various and develop rapidly. This paper introduces briefly the principles of classification and conformity assessment for IVD medical that proposed by GHTF (Global Harmonization Task Force).


Subject(s)
Diagnostic Techniques and Procedures , Equipment and Supplies, Hospital , Classification
13.
Article in Chinese | WPRIM | ID: wpr-407212

ABSTRACT

BACKGROUND: Is the inhibition of the hematopoietic stem progenitor cell (HSPC) proliferation and differentiation after human cytomegalovirus (HCMV) infection associated with abnormal expression of infected cell proliferated gene?OBJECTIVE: To observe the HOXB6-mRNA expression in the process of proliferation and differentiation of HCMV-infected HSPC into colony-forming unit granulocyte-macrophage (CFU-GM) and colony-forming unit erythroid (CFU-E).DESIGN: A controlled observation.SETTING: Laboratory for Molecular Biology, Affiliated Hospital of Luzhou Medical College, Lanzhou, Gansu Province, China.MATERIALS: All cord blood (CB) specimens were provided by the Obstetrics Department of Affiliated Hospital of Luzhon Medical College. They were collected from the umbilical vein of normal term neonates delivered spontaneously. All neonate mothers were healthy and HBS-Ag-negative. HCMV-IgM antibody revealed by routine ELUSA and HCMV-DNA checked by PCR were undetectable. Written informed consent for the laboratory measurements was obtained from each neonate mother, and the protocol was approved by the hospital's Ethics Committee. HCMV-AD169 strains were obtained from the Institute of Virology, Chinese Academy of Preventive Medicine. All-trans retinoic acid (ATRA, lot No. 20010126) was provided by Chongqing Huapont Pharm. Co., Ltd., China.METHODS: This study was performed at the Laboratory of Molecular Biology (state-level), Affiliated Hospital of Luzhou Medical College of Luzhou Medical College from April 2006 to April 2007. Cord blood mononuclear cells were separated for HSPC culture. According to different interventions, the study consisted of 4 groups. Control group: no HCMV virus solution was added and equal volume of culture medium was added instead. HCMV group: 105 PFU/mL HCMV-AD169 virus solution was added to the culture system. ATRA group: ATRA was added into the cultivation system at the final concentration of 60 μ mol/L. HCMV+ATRA group: ATRA was added into the HCMV group, and its final concentration was also 60 μ mol/L.MAIN OUTCOME MEASURES: In each group, cells were harvested on days 3,7 and 12. HOXB6 mRNA expression levels in CFU-GM and CFU-E were detected by real-time fluorescent-based quantification PCR.RESULTS: In the control group, both CFU-E and CFU-GM expressed HOXB6-mRNA. The HOXB6 mRNA expression was increased as a function of time. The HOXB6-mRNA expressed by CFU-E reached its peak level on day 12, while that expressed by CFU-GM reached its peak level on day 7. Compared to control group, the expression levels of CFU-E and CFU-GM HOXB6-mRNA genes in normal cord blood were significantly lower in the HCMV group (P<0.05)and significantly higher in the ATRA group (P<0.05) at each time point after HCMV infection. Furthermore, the expression levels were significantly higher in the ATRA+HCMV group than in the HCMV group at each time point(P<0.05-0.01).CONCLUSION: HOXB6-mRNA expression is stable and lasting in the proliferation and differentiation of HSPC into CFU-GM and CFU-E. HCMV could down regulate HOXB6 gene expression, and ATRA could up regulate HOXB6 gene expression.

14.
Article in Chinese | WPRIM | ID: wpr-587992

ABSTRACT

Objective To evaluate the short-term efficacy of the use of Y-shaped mesh in the treatment of anterior vaginal wall cystocele and stress urinary incontinence. Methods A total of 34 women diagnosed as having anterior vaginal wall cystocele complicated with stress urinary incontinence underwent intravaginal slingplasty using a Y-shaped mesh from April 2003 to May 2006. Evaluations on symptom improvement and postoperative recurrence rate were made. Results All the patients could spontaneously void urine after catheter removal at 24 hours after operation, with the residual urine less than 100 ml. Follow-up checkups for 3~37 months (mean, 26 months) found no urinary retention, urinary tract infection, bladder dysfunction, or recurrence. Conclusions Use of Y-shaped mesh in the treatment of both anterior vaginal wall cystocele and strees urinary incontinence is feasible.

15.
Article in Chinese | WPRIM | ID: wpr-280959

ABSTRACT

<p><b>OBJECTIVE</b>To develop a new nonisotopic detection method of enzyme-amplified time-resolved fluorescence (EATRF) or enzyme-amplified lanthanide luminescence (EALL) for nucleic acid hybridization assays, which can be applied extensively in clinical diagnosis.</p><p><b>METHODS</b>The method combines the high affinity of biotin-streptavidin system, amplification of enzyme, and inherent advantage of lanthanide chelate with the background elimination of time-resolved fluorescence detection. The conversion of 5-fluorosalicyl phosphate to 5-fluorosalicylic acid (5-FSA) by alkaline phosphatase. The salicylic acid product forms a luminescent ternary chelate with Tb3+ and EDTA.</p><p><b>RESULTS</b>The dynamic range of the standard curve of EATRFA for nucleic acid hybridization assay was very wide, the range was more than third order of magnitude. The detection sensitivity was about 10 pg of target sequence. When the known target sequence was 20, 10 and 2 ng, the ratio of measured amount to known amount was 110%, 90% and 115% respectively. The main experimental conditions, for example, the irradiating time of ultraviolet rays, the concentrations of biotinylated probe, AP-SA, 5-FSAP and Tb-EDTA and the methods of washing in the related steps, have been optimized. A new stable technology of fluorescence has been developted.</p><p><b>CONCLUSIONS</b>EATRF detection for nucleic acid hybridization assays is a new sensitive simple method, which has a great prospect.</p>


Subject(s)
Alkaline Phosphatase , Blotting, Southern , DNA , Genetics , Fluoroimmunoassay , Methods , Luminescent Measurements , Metals, Rare Earth , Nucleic Acid Hybridization , Methods , Spectrometry, Fluorescence , Substrate Specificity
16.
Article in Chinese | WPRIM | ID: wpr-640377

ABSTRACT

Objective To observe the expression level of HOXB6 gene on the differentiation and proliferation of hematopoietic stem cells(HSC) to colony forming unit-granulocyte-monocyte(CFU-GM),erythroid progenitor(CFU-E) and colony forming unite-T-lymphocyt(CFU-TL).And the proliferation procress was affected by all-transretinoic acid(ATRA).Methods 1.By the colony culture in vitro,the impact of ATRA on the CFU-GM,CFU-E,CFU-TL colony formation were surveyed;the expressions of HOXB6 gene were observed on the differentiation procress of HSC to CFU-GM,CFU-E and CFU-TL affected by ATRA on the 3rd,7th,12th.2.Real-time fluorogenic quantitative reserve transcription-polymerize chain reaction(FQ-RT-PCR) method was used to explore the possible mechanism of ATRA up-regulated to human cord blood CFU-GM,CFU-E and CFU-TL in gentic level.Results 1.HOXB6 gene had a regulatory function in the differentiation procress of hematopoiesis.During the differentiation and proliferation of HSC to colony forming CFU-GM or CFU-E,CFU-TL in vitro,the expressions of HOXB6 gene were significant positive.2.Compared with the expressions of CFU-GM and CFU-TL HOXB6 mRNA on day 3,the quantity of HOXB6 mRNA was obviously higher on day 7 and lower on day 12,respectively in each group.Compared with the expression of CFU-E HOXB6 mRNA on day 3,the quantity of HOXB6 mRNA was obviously higher on day 12 in each group.3.Compared with the HOXB6 gene of control group,the expressions of HOXB6 gene of group ATRA were up-regulated remarkable.Conclusions 1.HOXB6 gene has a regulatory function in the differentiation procress of hematopoiesis.HOXB6 gene plays an important role in the progress which can be associated with the regulating effect of HOXB6 gene on the differentiation and proliferation of CFU-GM,CFU-E and CFU-TL.2.During the differentiation and proliferation of HSC,the expression of HOXB6 gene is regular in time.3.Low concentration of ATRA(60 ?mol?L-1) can up-regulate the expression of HOXB6 gene,which confirms the theory that the normal hematopoie-tic lineage determination and maturation rely on the stable and consistent expression of HOXB6 gene.

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