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Objective:To learn about the awareness, education status and willingness of iodine deficiency disorders (IDD) among elementary school students in Guizhou Province, and to provide a scientific basis for exploring intervention strategies for health education of iodine deficiency in the future.Methods:From June 2021 to May 2022, each IDD monitoring county was selected from the east, south, west, north and middle directions of Guizhou Province, and one elementary school was selected from each county. All students in two classes of Grade 5 and Grade 6 were selected in whole groups to conduct on-site questionnaire surveys in the form of anonymous examinations. The survey mainly included general demographic information and IDD awareness, education status and willingness, and binary logistic regression was used to analyze the relevant influencing factors.Results:A total of 1 259 elementary school students in Guizhou Province were investigated, the rates of awareness of IDD, acceptance of IDD publicity and education, and willingness to accept IDD publicity and education among elementary school students were 37.7% (1 900/5 036), 25.1% (316/1 259) and 69.6% (876/1 259), respectively. By binary logistic regression analysis, gender, residence, grade and father's education level were the influencing factors of pupils' awareness of iodine deficiency ( P < 0.05); residence, age and father's education level were the influential factors of elementary school students receiving iodine deficiency education ( P < 0.05); gender, residence, ethnicity and whether the child was the only child or not were the influential factors of elementary school students' willingness to accept IDD education ( P < 0.05). Conclusions:The elementary school students in Guizhou Province have insufficient knowledge about IDD. The publicity and education for iodine deficiency prevention is limited, and the students' willingness to learn is not high. The publicity, education and intervention for iodine deficiency prevention among elementary school students should be comprehensively strengthened.
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BACKGROUND:At present, mouse embryonic neural stem cells (NSCs) culture has been skillfully operated by many labs, but there are differences existing about which part are dissociated to get NSCs. Embryonic 14 days (E14) mouse brain tissues are widely used for culturing NSCs, but there are less studies about the detailed percentage and difference of NSCs separated from different brain tissues. OBJECTIVE: To test the proportion and difference of NSCs and neurons percentage from E14 mouse whole brain, cortex and forebrain, providing quantized data for optimizing the isolation of high-purity NSCs. METHODS:E14 C57BL/6 mouse whole brain, cortex and forebrain tissues were separated and dissociated into single cells that were adherently cultured for 3.0-4.0 hours and labeled by DAPI. Then the cells were immunostained with NSCs specific marker, Nestin, and neuron specific marker, Tuj1, to identify NSCs and neurons percentage by calculating Nestin+/DAPI and Tuj1+/DAPI. In addition, real-time PCR assay was used to test Nestin and Tuj1 mRNA expression in the E14 mouse whole brain, cortex and forebrain. RESULTS AND CONCLUSION: (1) Immunocytochemical results showed that there were a large amount of Nestin+ and Tuj1+ cells in the whole brain, cortex and forebrain of E14 mice. NSCs percentage in the forebrain was obviously higher than that in the whole brain (P 0.05); the Tuj1 mRNA expression in the forebrain was significantly lower than that in the whole brain (P < 0.05) and in the cortex (P < 0.05). These findings indicated that the forebrain had the most NSCs and the least neurons compared with the whole brain and the cortex. In summary, E14 mouse forebrain has the highest percentage of NSCs compared with the whole brain and cortex, which is a better source to obtain NSCs for the following cell culture experiments.
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Objective To understand and compare the proportion of neural stem cells (NSCs) in the whole brain and cerebral cortex of mice at different embryonic days, and provide quantitative data for the later optimization of NSCs isolation and culture.Methods The whole brains (at embryonic 12.5, 14, 16 and 18 days) and cerebral cortex (at embryonic14, 16 and 18 days) were isolated and digested into single cell suspension, and were adherently cultured for 3-4 h.Immunofluorescence staining of Nestin, a NSCs specific marker, was used to statistically analyze the proportion of NSCs in each group.Expression of Nestin mRNA in the cerebral cortex of mice at E12.5, E14, E16, and E18 was detected by real-time fluorescence quantitative PCR.Results The result of immunofluorescence assay showed that there were Nestin-positive cells in the whole brain and cerebral cortex of mice at different embryonic days.In the whole brain,the proportion of NSCs was highest at E12.5 (53.42±1.57%) and lowest at E18(25.96±1.31%), and the proportions at E14 and E16 were placed in the middle among the groups.In the cerebral cortex, the highest proportion of NSCs was at E14 (33.65±0.29%), and the lowest at E18(25.29±0.28%), and the middle at E16 (26.82±0.30%).The result of real-time PCR showed that when the mRNA expression of Nestin in the cerebral cortex was set to 1, the relative mRNA expression of Nestin was 0.83±0.04 at E14, 0.77±0.05 at E16, and 0.44 ±0.05 at E18.Thus, the mRNA expression level of Nestin in the mouse cerebral cortex was gradually decreasing with the increase of embryonic days.Conclusions During the brain development, the proportion of NSCs is gradually decreasing in the whole brain and cerebral cortex of mice with the increase of embryonic days.
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Objective To study the effects of different differentiation times (one,3 and 5 d) on mouse embryonic neural stem cells (NSCs) differentiating into astrocytes and neurons.Methods (1) Fetal cortices of embryonic 14 d (El4) C57BL/6 mice were isolated,digested and cultured.The nestin and Sox2 expressions in the second passaged neural spheres and monolayer cultured NSCs were detected by immunofluorescent staining.(2) And then second passaged neural spheres were digested into NSCs;they were inoculated in the slide and overnight cultured,and then,they were changed into the differential medium the next day;immunofluorescence assay was used to observe the glial fibrillary acidic protein (GFAP) and Tujl expressions;the averaged numbers,mean longest length and mean branches of the neuritis were analyzed one,3 and 5 d after differentiation.(3) Real time fluorogenic quantitative PCR was used to detect the nestin,GFAP and Tuj1 mRNA expressions one,3 and 5 d after differentiation of NSCs.Results (1) NSCs were successfully cultured and almost of all cells were both nestin and Sox2 positive NSCs.(2) Immunofluorescence assay showed that as the differentiation time increasing,numbers of differentiated astrocytes and neurons became larger,and their morphologies became more complicated.The cell counting results showed that:as compared with one d group,3 and 5 d groups had significantly high GFAP+ astrocyte percentages (P<0.05);as compared with one and 3 d groups,5 d group had significantly higher Tuj 1+ neuronal percentage (P<0.05);as compared with one d group,3 and 5 d groups had significantly larger averaged neurite numbers (/P<0.05);the averaged longest neurite length was increased as differentiation time increasing and there was obvious difference between each two time points (P<0.05);as compared with one d group,3 and 5 d groups had significantly higher averaged neurite branching levels (P<0.05).(3) And PCR results mainly showed that nestin mRNA expression in one d group was significantly higher than that in 3 and 5 d groups (P<0.05);GFAP and Tuj1 mRNA expressions in 5 d group were statistically higher than those in one and 3 d groups (P<0.05).Conclusion With the differentiation time increasing,the numbers of differentiated astrocytes and neurons become large,their morphologies become complicated,and their percentages are gradually increased;more mature neurites are noted;nestin mRNA expression gradually decreases while GFAP and Tuj1 mRNA expressions gradually increase in monolayer cultured mouse embryonic NSCs.
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Objective To explore the resistance of neural stem cells (NSCs) in β-amyloid (Aβ) toxicity after being transfected with brain-derived neurotrophic factor (BDNF) and its mechenism.Methods NSCs in the C57 mice were transfected with BDNF gene by using lipofectamine technique; the BDNF gene expression in transfected NSCs and their differentiated cells were detected by immune-fluorescent staining and ELISA.Rat neurons were cultured in vitro and divided into 3 groups:neurons group,neurons co-cultured with NSCs group,neurons co-cultured with BDNF transfected NSCs group; two-parallel group design was adopted.Three d after the group culturing,cells from each parallel group were added Aβ40 protein.Neuron survival of each group was recorded 24 and 48 h after adding Aβ40 by ELISA.The Aβ,tau,and phosphorylating tau (ptau) levels were detected by Western blotting.Results BDNF-transfected NSCs could continuously express BDNF gene products during the first 1.5 weeks.Both transfected NSCs and their progenies expressed BDNF gene products.Using neurons group as reference,the mean survival rates of neurons plus Aβ40 group,neurons co-cultured with NSCs plus Aβ40 group,neurons co-cultured with BDNFtransfected NSCs plus Aβ40 group were 51.86%±0.03%,73.68%±0.04% and 85.34%±0.01% at 24 h after adding Aβ40,and 38.76 %±0.01%,59.65 %±0.04%,75.61%± 0.07% at 48 h after adding Aβ40; the survival rate in the neurons plus Aβ40 group and neurons co-cultured with NSCs plus Aβ40 group was significantly lower than the other groups (P<0.05).High level of Aβ protein was noted in the neurons plus Aβ40 group,neurons co-cultured with NSCs plus Aβ40 group and neurons co-cultured with BDNF transfected NSCs plus Aβ40 group,without significant differences between each two groups (P>0.05).The tau protein level showed no significant differences between each two groups (P>0.05),while the p-tau protein level was significantly different (that in the neurons co-cultured with NSCs plus Aβ40 group and neurons co-cultured with BDNF transfected NSCs plus Aβ40 group was significantly higher than that in the other groups,P<0.05).Conclusion BDNF-transfected NSCs express BDNF gene products during the early time,which alleviate the toxicity of Aβ to NSCs and reduce the p-tau protein level,leading to increased survival rate of neurons.
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Objective To reduce experimental costs and improve the utilization of S9, we use spiral coating technique to evaluate the activity of rat liver S9 prepared by combination inducing method as well as to establish cryopreservation method.Methods Using spiral coating technique and Ames test to evaluate the activity of self-made rat liver S9 and commercially available S9 separately.We use glycerinum as protective agent to establish cryogenic storage method, so that S9 can be in liquid form stored at -20 °C in the refrigerator.Results In the Ames assay as well as using spiral coating technique, the number of revertant colonies had dose-response relationship among the dose of S9.When conditions were the same, the number of revertant colonies in positive control was at the approximate level in presense of self-made rat liver S9 and commercially available S9 respectively.When S9 ( concentration of 38%) was added to the amount at 1.48 ~6.62 μL /μL broth dose of bacteria, it can significantly induced Salmonella typhimurium histidine strains TA100, reverse mutation rates were three times more than the control group.Conclusions Spiral coating technique can successfully evaluate the activity of rat liver S9.The inducing method of combination of PB and BF can take the place of the unducing method of PCBs in the preparation of Liver homogenate S9.
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Objective To explore the change of serum Apelin level and its relationship with insulin resistance (IR) in infertility patients with polycystic ovarian syndrome (PCOS). Methods Ninety-eight infertility patients with PCOS (PCOS group) and 72 infertility patients without PCOS (non-PCOS group) visiting our hospital for the first time from January 2011 to June 2012 were selected. The BMI , serum Apelin level (detected by ELISA), fasting blood glucose (FPG, detected by glucose oxidase method), fasting insulin (Fins, detected by chemiluminescence), and homeostasis model assessment of insulin resistance index (HOMA-IR) of the two groups were detected. Results The serum Apelin level and HOMA-IR in PCOS patients were higher than those in non-PCOS patients (3.28 ± 1.24 vs. 1.94 ± 0.78, P < 0.05; 3.84 ± 1.23 vs. 2.14 ± 0.77,P < 0.05). Pearson correlation analysis showed that serum Apelin level was positively correlated with HOMA-IR in PCOS patients (r=0.65, P<0.01). Conclusions The serum Apelin level in infertility patients with PCOS increased evidently, and was positively correlated with HOMA-IR; which indicated that Apelin may involve in the development of IR in infertility patients with PCOS.
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Objective To develop and evaluate the Quality of Life Instruments for Chronic Diseases -Osteoarthritis ( QLICD-OA). Methods The QLICD-OA was evaluated by testing 140 in-patients with Osteoarthritis. The statistical methods used included correlation analysis, factor analysis, paired t test and Generalizability coefficient. Results The rest-retest reliability γ of QLICD-OA was 0.91, Cronbach’s α for overall scale was 0.98 and Generalizability coefficient was 0.94. Correlation and factor analysis demonstrated good construct validity. After treatment, statistically significant scores were found in the tested group for Specific module and total instrument. Conclusions The QLICD-OA has good validity, reliability and responsiveness. The QLICD-OA may serve as the QOL measurement scale for patients with Osteoarthritis in China.
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The study intended to evaluate the combined examination of transient flash visual evoked potential (T-FVEP) and steady-state flash visual evoked potential (SS-FVEP) in diagnosis of optic neuropathy and macular disease. T-FVEP and SS-FVEP were examined in 22 cases (29 eyes) with optic neuropathy, 8 cases (9 eyes) with macular disease and 32 cases (32 eyes) of controls. The positive rates of T-FVEP and SS-FVEP in diagnosis of optic neuropathy were 83% and 100% respectively; that of combined examination was 100%. The positive rates of T-FVEP and SS-FVEP in diagnosis of macular disease were 5/9 and 7/9, and that of combined examination was 8/9. The positive rates of combined examination of T-FVEP and SS-FVEP in diagnosis of optic neuropathy and macular disease are higher than that of T-FVEP and SS-FVEP alone, and should be used routinely in the diagnosis of visual pathway disease.
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Objective To discuss the clinical value of simultaneous morcellation in situ in laparoscopic myomectomy (LM). Methods One hundred and eighty-nine women who underwent LM for symptomatic myomas were randomly divided into in situ group(94 cases) and standard group(95 cases). The operation time, hemorrhage perioperative,exhaust time postoperative, length of hospital stay were compared.Results The operation was successful in two groups. The operation time and hemorrhage perioperative in situ group were significantly less than those in standard group [ (98.20 ± 40.70) min vs. ( 129.20 ± 68.50)min (P<0.05) and (113.40± 38.30) ml vs. (168.90±61.70) ml (P<0.01)]. There was no statistical difference in exhaust time postoperative, length of hospital stay between two groups (P >0.05). One case occurred eechymoma in two groups respectively. Conclusion The application of simultaneous morcellation in situ in LM is safe and feasible.
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Objective To investigate the relationship of the chemical coding enteric nervous system of the mice and expression for neurotransmittor of enteric primary afferent neurons for Nociceptors.Methods Immunocytochemical and morphometric techniques were used to quantify the distribution of IB4-containing neurons in mice enteric nervous system using three mice chiocing every vision 50 neurons undering confocal microscopy IB4 immunolabelling and colocalized with calretinin and lectin B4.Results IB4 being binded to primary afferent neurons of enteric pleuxes happend in small intestin and colon of mice,where it was selective for nociceptive neurons.IB4 revealed large round or oval(Dogiel type II)neurons,type I neurons with prominent laminar dendrites and small neurons of myenteric ganglia.The type II neurons were immunoreactive for calretinin,and some type I neurons were immunoreactive for nitric oxide synthase.Most neurons in the submucosal ganglia bound IB4,and some of these were vasoactive intestinal peptide immunoreactive.Conclusion The results indicate that IB4 labels specific subgroups of enteric neurons in the enteric nervous system of the mice.These include intrinsic primary afferent neurons,but other neurons,including secretomotor neurons,are labeled.The results suggest that IB4 is not a specific label for enteric nociceptive neurons.
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dbcAMP was injected i. p. into four U_(14) ascitic tumor bearing mice on 3,4,5,6 and 7 days after inoculation of tumor cells. The ascites was aspirated 0.5-1 hour after last injection. For scanning electron microscopy, the tumor cells, washed twice in Hanks solution, were fixed in glutaraldehyde and OsO_4, dehydrated with ethyl alcohol and dried with camphene. For flow microcytometric analysis, the treated tumor cells were fixed in 70% cold ethyl alcohol and stained with propidium iodide. Under SEM, the untreated tumor cells were large, spherical, and uniform in size with numerous long thin microvilli on the cell surface. A few barely visible minute protrusions were present on the microvilli and cell membrane. The great majority of treated tumor cells became smaller and variable in size, with shortened microvilli which reduced in number and even disappeared in some area. Many granular protrusions, larger than that of the control, were clearly observed on the cell membrane and microvilli. The result of flow microcytometric analysis showed that the DNA histogram and cell cycle profile from dbcAMP treated cells have no significant difference from the untreated controls, so it is evident that morphologic changes resulted from dbcAMP were not caused by cell cycle alteration. The morphogenesis of microvilli and cell membrane changes in dbcAMP treated cells is not clear. The relation of these configurations to differentiation of malignant tumor cells is discussed.
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Our study demonstrated that ARS cells can be induced to differentiate into macrophage-like cells with changes in phenotype, nonspecific esterase activity and phagocytic function by adding ginsenosides in short-term cultures. Synthesis of DNA, mitosis and the growth of the culture cells transplanted in mouse are also inhibited in this condition. The size of cells, nuclei and nucleoli, as well as nuclear-cytoplasmic ratio of ginsenosides treated cells are diminished significantly. Microvilli of these cells are reduced in number with formation of. ruffles on the cell surface. Mitochondria are increased, their size and distribution become regular. The fact that numerous small cells, induced by ginsenosides exhibit the most conspicuous alteration mentioned above along with marked phagocytic activity indicates that they are highly differentiated macrophage-like cells. Whether the inhibition of cell growth and induction of differentiation by ginsenosides is caused through its action on the molecules regulating the gene expression of cell growth and differentiation needs further study.
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Murine ARS reticulum sarcoma cells, when incubated with 1.5, 2.0 and 2.5% DMSO in vitro, were induced to differentiate into macrophage like cells. They showed reduction of cellular and nuclear size and lowering of nucleo-cytoplasmic ratio with marked change in nuclear size. The most effective one was 2.5% DMSO which caused marked reduction of nucleolar size. Tumor cells with nucleo-cytoplasmic ratio less than 0.5 and with reduced nucleoli resembled macrophage. 2.5% DMSO induced the increase of non-specific esterase positive cells and the increase of polystyrene phagocytic percentage and phagocytic index. Under EM, 2.5% DMSO treated ARS cells with nucleocytoplasmic ratio less than 0.5 exhibited abundant cytoplasmic organelles, developed Golgi complex and secondary lysosomes. Scanning EM showed that many large cells with numerous dense slender microvilli were observed in the control. Under the action of 2.5% DMSO, large cells were reduced, small cells were increased and microvilli markedly decreased and shortened. These small cells appeared to be differentiated macrophage like cells and were similar to small cells seen under EM and light microscope. In 2.5% DMSO treated groups, the mitotic index of ARS cells were markedly lowered.