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AIM: To assess the repeatability and agreement of higher-order aberration obtained by adaptive optics visual simulator(VAO)compared with OPD-Scan Ⅲ.METHODS: A cross-sectional study was conducted from August to September 2023, including a total of 204 patients(204 eyes)with myopia whose right eyes were measured. The examinations were performed by the same skilled examiner using both devices separately. The VAO device was used to measure higher order aberrations of orders 3 to 6 at a pupil diameter of 4.5 mm, while both the VAO and OPD-Scan Ⅲ devices were utilized to measure total higher-order aberration(tHOA), spherical aberration(SA), coma aberration(Coma), and trefoil aberration(Trefoil)of the entire eye at pupil diameters ranging from 3 to 6 mm. Furthermore, the repeatability of whole eye aberration measurements obtained with the VAO device was evaluated and the agreement of the two devices was assessed.RESULTS: The whole-eye higher-order aberrations measured by VAO demonstrated excellent repeatability(0.767≤ICC≤0.941, Sw<0.01 μm, TRT<0.1 μm). There was no statistically significant difference in Coma measured by VAO or OPD-Scan Ⅲ for pupil diameters ranging from 4 to 6 mm(P>0.05), while a statistically significant difference was observed in whole-eye tHOA of other pupil diameters(all P<0.05). The agreement of aberration measurements for each order between VAO and OPD-Scan Ⅲ for 3 mm pupil diameters, SA at 4 and 5 mm pupil diameter and Coma at 4 mm pupil diameter showed a 95% limit of agreement(LoA)<0.1, indicating good agreement; however, poor agreement was found for the remaining aberration measurements at different pupil diameters, with a 95%LoA>0.1, and there were significant differences in higher-order aberrations measured by two devices under a pupil diameter of 3 mm(r=0.218-0.317, P<0.01), 4 mm(r=0.406-0.672, P<0.01), 5 mm(r=0.538-0.839, P<0.01 and r=0.030-0.109, P>0.01)and 6 mm(r=0.369-0.766, P<0.01).CONCLUSION: The VAO demonstrates favorable repeatability when assessing whole-eye higher order aberration under pupil diameters of 3-6 mm. However, there is inadequate agreement and interchangeability in whole-eye higher order aberration at 3-6 mm pupil diameter between VAO and OPD-Scan Ⅲ for clinical purposes.
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Background/Aims@#Exacerbating factors of ulcerative colitis (UC) are multiple and complex with individual influence. We aimed to evaluate the efficacy of disease control by searching and restricting inflammation trigger factors of UC relapse individually in daily clinical practice. @*Methods@#Both patients with UC history or new diagnosis were asked to avoid dairy products at first doctor visit. Individual-reported potential trigger factors were restricted when UC flared up (Mayo endoscopy score ≥1) from remission status. The remission rate, duration to remission and medication were analyzed between the groups of factor restriction complete, incomplete and unknown. @*Results@#The total remission rate was 91.7% of 108 patients with complete restriction of dairy product. The duration to remission of UC history group was significantly longer than that of new diagnosis group (88.5 days vs. 43.4 days, P=0.006) in patients with initial endoscopic score 2–3, but no difference in patients with score 1. After first remission, the inflammation trigger factors in 161 relapse episodes of 72 patients were multiple and personal. Milk/dairy products, herb medicine/Chinese tonic food and dietary supplement were the common factors, followed by psychological issues, non-dietary factors (smoking cessation, cosmetic products) and discontinuation of medication by patients themselves. Factor unknown accounted for 14.1% of patients. The benefits of factor complete restriction included shorter duration to remission (P<0.001), less steroid and biological agent use (P=0.022) when compared to incomplete restriction or factor unknown group. @*Conclusions@#Restriction of dairy diet first then searching and restricting trigger factors personally if UC relapse can improve the disease control and downgrade the medication usage of UC patients in daily clinical practice.
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OBJECTIVE To establish and apply a method for simultaneous determination of the contents of dicentrine, protopine and coptisine in Tibetan Corydalis pallida of different origins, and to provide reference for origin determination and quality control of the kind of medicinal materials. METHODS Ultra performance liquid chromatography-tandem quadrupole mass spectrometry method was used. The determination was performed on Agilent EC-C18 column (100 mm×2.1 mm, 2.7 μm) with mobile phase consisted of acetonitrile-0.1% formic acid by gradient elution. The flow rate was 0.2 mL/min, and the column temperature was set at 35 ℃ . MS detection was carried out by electrospray ionization in positive modes, multiple reaction monitoring mode was used for quantitative analysis. RESULTS The injection mass concentrations of dicentrine, protopine, coptisine ranged from 5.88 to 117.60, 53.70 to 1 074.00, and 4.85 to 97.00 ng/mL, respectively, showing a good linear relationship with their respective peak areas (r=0.998 2, 0.991 9, and 0.999 6, respectively). The limits of quantitation were 2.35, 1.07 and 1.46 ng/mL; the limits of detection were 1.17, 0.54, 0.49 ng/mL, respectively. RSDs of precision, stability (24 h) and repeatability tests were all lower than 2.0%. The average recovery rates were 97.41%, 98.89% and 105.44%( all RSDs<5.0%, n=6). CONCLUSIONS The established method has good selectivity and high accuracy, and is suitable for the rapid analysis of dicentrine, protopine and coptisine in Corydalis. The total contents of three alkaloids in different original medicinal materials are from high to low in order of C. chrysosphaera, C. mucronifera, C. pygmaea, C. hendersonii and C. conspersa. The alkaloid contents in C. chrysosphaera and C. mucronifera are relatively similar, but no dicentrine has been detected in C. conspersa and C. hendersonii.
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Objective:To investigate the inhibitory effect of curcumin on oxidative stress in BCG-infected macrophages based on the Nrf2 pathway.Methods:THP-1-derived macrophages were infected.The experiment was divided into control group,BCG group,BCG+curcumin group and BCG+curcumin+ML385 group.Cellular ROS fluorescence intensity were observed under a fluores-cence microscope;Glutathione(GSH)levels were measured by Colorimetry;Western blot was used to detect the protein expressions of Nrf2,HO-1 and NQO1;MTT was used to detect the proliferation rate of macrophages.Results:BCG infection significantly enhanced ROS fluorescence intensity,reduced cell GSH content(P<0.01),inhibited protein expressions of Nrf2,HO-1 and NQO1,at the same time inhibited cell proliferation(P<0.01);curcumin significantly weakened ROS fluorescence intensity,increased GSH level(P<0.05),promoted Nrf2,HO-1 and NQO1 protein expressions and cell proliferation(P<0.01);Nrf2 inhibitor ML385 reversed the effect of curcumin.Conclusion:Curcumin can alleviate BCG-induced oxidative stress in macrophages by increasing the expression of Nrf2 and inducing the transcription of downstream antioxidant molecules.
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Objective:To investigate the diagnostic value of a single hydrogen-methane breath test (SHMBT) for small intestinal bacterial overgrowth (SIBO).Method:The current investigation was a cross-sectional study. Questionnaires and SHMBTs were administered to 162 patients with gastrointestinal symptoms (case group) and 69 healthy volunteers (control group). Differences in SHMBT results between the two groups were assessed,and cut-off values of CH 4 (methane) and H 2 (hydrogen) were analyzed via receiver operating characteristic (ROC) curves. Lastly,archived SHMBT data from 2 655 patients with gastrointestinal symptoms (validation set) were used to evaluate the diagnostic value of the SHMBT with respect to SIBO. The Chi-square test,the Mann-Whitney U test,Spearman′s Rank correlation analysis,and the Z test were used for statistical analysis. Results:Based on the international recommended diagnostic criteria for SIBO,which are fasting CH 4 ≥10 ppm (parts per million) or H 2 ≥20 ppm,the SHMBT-positive rate in the case group was significantly higher than that of control group (35.2% vs. 21.7%, χ2=4.08, P=0.043). Levels of CH 4 and H 2 were higher in the case group than in the control group [CH 4: 3(2,7) vs. 3(1,3) ppm, H 2: 11(4,22) vs. 10(5,15) ppm],and the difference in CH 4 levels was statistically significant ( Z=6.22, P=0.001). ROC curves were generated based on whether the subjects had gastrointestinal symptoms. The areas under the ROC curves were 0.633 for CH 4 alone,0.531 for H 2 alone, and 0.620 for CH 4 combined with H 2. The cut-off values were fasting CH 4≥4 ppm,fasting H 2≥13 ppm,and fasting CH 4 ≥5 ppm (or CH 4≥4 ppm and H 2≥24 ppm),respectively. Measuring CH 4 alone and CH 4 combined with H 2 was effective for determining the presence of gastrointestinal symptoms ( P<0.05). When CH 4 alone or CH 4 combined with H 2 were used as diagnostic indicators of SIBO, the respective SHMBT-positive rates in the validation set were 34.2% and 30.4%. These rates did not significantly differ from the SIBO-positive rate of 32.0% obtained via the international recommended diagnostic criteria ( P>0.05). The specificity of CH 4 alone was 79.9%,and the accuracy of CH 4 alone was 68.8%. The specificity of CH 4 combined with H 2 was 85.0%,and the accuracy of CH 4 combined with H 2 was 71.7%. Conclusion:Rapid one-time determination of CH 4 and H 2 in exhaled breath may a viable diagnostic method for SIBO, and using CH 4 combined with H 2 ( i.e.,fasting CH 4≥5 ppm, or CH 4 ≥4 ppm and H 2 ≥24 ppm) as cutoff values may be feasible.
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Genetic robustness refers to the fact that an organism has a buffer system that can maintain normal development, even if the existence of genetic mutations during biological development. Previous research on the underlying mechanism of genetic robustness mainly involves in genetic redundancy and distributed robustness, both of which are triggered at the protein level. Recently, a new genetic compensation response (GCR) mechanism has been discovered in zebrafish, which occurs in knockout rather than knockdown individuals and is triggered upstream of protein feedback regulation. Since there are many concepts related to genetic robustness, this review attempts to clarify these concepts from the types of compensation genes and triggering modes. Additionally, we aim to understand the latest discovered GCR mechanism and look forward to studying the function of specific genes based on functional compensation mechanisms.
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Objective:To establish the normal reference range of the ratio of fetal umbilical venous flow rate to umbilical artery pulsatility index (VAI).Methods:A total of 816 normal fetuses underwent prenatal examination and delivery were randomly selected from October 2018 to December 2020 in Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region. Fetal weight was obtained by measuring fetal biparietal diameter, head circumference, abdominal circumference, and femoral length.Umbilical venous flow (Quv) was measured. Umbilical artery pulsatility index (UA-PI) was obtained in the free segment of amniotic fluid. Quv was standardized according to fetal size to calculate the umbilical venous flow rate (nQuv) and VAI. The association between Quv, nQuv, UA-PI, VAI and the fetal gestational week were analyzed using correlation analysis. VAI was presented as ± s, the upper limit of 95% reference value and the lower limit of 5% reference value were taken as the standards of VAI increase and decrease, respectively. Twenty-six fetuses whose VAI were lower than limit of 5% and 20 fetuse whose VAI were than limit of 95% were chosed as the case group. Results:①Fetal Quv was positively correlated with gestational week ( r=0.893, P<0.001), nQuv and UA-PI were negatively correlated with gestational week ( r=-0.552, -0.827; all P<0.001), and VAI had no significant correlation with gestational week ( r=0.000, P=0.758); ②The mean, standard deviation, lower 5% reference value, and upper 95% reference value of VAI were 195.81, 55.61, 105.95, and 293.33, respectively; ③In the cases with abnormal VAI, 26 fetuses with reduced VAI, of whom there were 16 cases of maternal hypertension, and 13 cases complicated by severe preeclampsia; 1 case with 40 turns of umbilical cord torsion, 3 cases of stillbirth, 16 cases of preterm delivery, 19 cases of low neonatal birth body weight, 4 cases of 1-min Apgar score ≤7, 6 cases of umbilical artery blood pH<7.2, and 1 case without abnormalities in fetus during pregnancy and follow-up newborn. Among the 20 fetuses with increased VAI, there were 10 cases of fetal severe thalassemia, 2 cases of thalassemia, 1 case of sacrococcygeal teratoma, 1 case of portal venous shunt, 3 cases of placental chorioangioma, and 3 cases without abnormalities in fetus during pregnancy and follow-up newborn. Conclusions:The measurement and calculation of fetal VAI is simple and easy to perform. As a comprehensive index, fetal VAI remains constant in mid and late pregnancy, facilitates the follow-up of abnormal fetuses, and has potential clinical application.
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Objective:To study the effect of rehabilitation training on the expression of neuroglobin (Ngb), oxidative stress and axon regeneration in the cortex and explore possible mechanisms of functional recovery after cerebral infarction.Methods:Thirty-six male Sprague-Dawley rats were randomly divided into a sham operation group, a model group and a rehabilitation group. Cerebral infarction was modelled in the model and rehabilitation groups using Longa′s middle cerebral artery occlusion (MCAO) technique. The sham operation group received the same procedure except that no thread was inserted to block the middle cerebral artery. The rats in the rehabilitation group began treadmill training 24h after the operation, while the other two groups were left on the treadmill without training. On the 3rd, 7th and 14th days after the operation, all of the rats′ neurological functioning was assessed using modified neurological severity scores (mNSSs). After the last mNSS test, all of the rats were sacrificed and peri-infarct brain tissue was resected to detect the expression of Ngb and oxidative stress indicators including superoxide dismutase (SOD), nitric oxide and malondialdehyde (MDA), as well as neurofilament-200 (NF-200) indicating axon regeneration.Results:On the 3rd day after the surgery there was no significant difference between the average mNSS scores of the rehabilitation and model groups. On the 7th and 14th day the average mNSS score of the rehabilitation group was significantly better than that of the model group. The average expression of Ngb in the model group was significantly higher than in the sham operation group. After the intervention, the average expression of SOD in the rehabilitation group was significantly higher than in the model group, while NO and MDA expression were significantly lower. After the intervention the average expression of NF-200 in the rehabilitation group was also significantly higher than in the model group.Conclusions:Rehabilitation training benefits the recovery of neurological function after cerebral infarction, at least in rats. The mechanism may be related to the upregulation of Ngb, alleviation of oxidative stress and enhancement of axonal regeneration in the peri-infarct cortex.
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OBJECTIVE@#To explore the differences between hematological phenotypes of patients with different genotypes in gene mutations and deletion α- thalassemia.@*METHODS@#By screening the α- thalassemia gene test results in the First Affiliated Hospital, Sun Yat-Sen University from January 2015 to April 2020, the patients with mutation and deletion α- thalassemia were obtained, then the differences between hematological phenotypes of patients with different genotypes were analyzed.@*RESULTS@#There were 96 patients with mutation combined with deletion α- thalassemia from the results of 24 054 α- thalassemia patients screened out, including 79 patients with non-deletion Hb H disease (α@*CONCLUSION@#The hematological phenotype changes caused by α
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Humans , Genotype , Mutation , Phenotype , Retrospective Studies , alpha-Thalassemia/geneticsABSTRACT
OBJECTIVES@#To study the expression levels of microRNA-138 (miR-138) and Runt-related transcription factor 3 (RUNX3) in peripheral blood of children with cough variant asthma (CVA) and their regulatory effects on Th1/Th2 balance.@*METHODS@#Sixty-five children with CVA (CVA group) and 30 healthy children (control group) were enrolled. Peripheral venous blood samples were collected for both groups, and CD4@*RESULTS@#Compared with the control group, the CVA group showed significantly decreased levels of IFN-γ and IL-2 from CD4@*CONCLUSIONS@#MiR-138 regulates Th1/Th2 balance by targeting RUNX3 in children with CVA, providing a new direction for the treatment of CVA.
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Child , Humans , Asthma , Core Binding Factor Alpha 3 Subunit/genetics , Cough , Interleukin-13 , MicroRNAs/genetics , Th1 Cells , Th1-Th2 Balance , Th2 CellsABSTRACT
BACKGROUND@#It is crucial to differentiate accurately glioma recurrence and pseudoprogression which have entirely different prognosis and require different treatment strategies. This study aimed to assess the diagnostic accuracy of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) as a tool for distinguishing glioma recurrence and pseudoprogression.@*METHODS@#According to particular criteria of inclusion and exclusion, related studies up to May 1, 2019, were thoroughly searched from several databases including PubMed, Embase, Cochrane Library, and Chinese biomedical databases. The quality assessment of diagnostic accuracy studies was applied to evaluate the quality of the included studies. By using the "mada" package in R, the heterogeneity, overall sensitivity, specificity, and diagnostic odds ratio were calculated. Moreover, funnel plots were used to visualize and estimate the publication bias in this study. The area under the summary receiver operating characteristic (SROC) curve was computed to display the diagnostic efficiency of DCE-MRI.@*RESULTS@#In the present meta-analysis, a total of 11 studies covering 616 patients were included. The results showed that the pooled sensitivity, specificity, and diagnostic odds ratio were 0.792 (95% confidence interval [CI] 0.707-0.857), 0.779 (95% CI 0.715-0.832), and 16.219 (97.5% CI 9.123-28.833), respectively. The value of the area under the SROC curve was 0.846. In addition, the SROC curve showed high sensitivities (>0.6) and low false positive rates (<0.5) from most of the included studies, which suggest that the results of our study were reliable. Furthermore, the funnel plot suggested the existence of publication bias.@*CONCLUSIONS@#While the DCE-MRI is not the perfect diagnostic tool for distinguishing glioma recurrence and pseudoprogression, it was capable of improving diagnostic accuracy. Hence, further investigations combining DCE-MRI with other imaging modalities are required to establish an efficient diagnostic method for glioma patients.
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Humans , Glioma/diagnostic imaging , Magnetic Resonance Imaging , Neoplasm Recurrence, Local/diagnostic imaging , ROC Curve , Sensitivity and SpecificityABSTRACT
Objective:To investigate the influence of long non-coding RNA-CRNDE in stem cell properties and their mechanism in glioma.Methods:The CD133 +U251 stem cells (U251s) were isolated from human glioma cell line U251 by immunomagnetic beads. Cells were divided into U251s group, U251s-CRNDE group, U251 group, and U251-CRNDE group; CRNDE shRNA lentiviral vectors were transfected into cells in the U251s-CRNDE group and U251-CRNDE group to down-regulate the CRNDE expression. The CRNDE mRNA expression was detected by real-time fluorescent quantitative PCR (RT-qPCR); CCK-8 assay was used to detect the cell viability; Transwell assay was used to detect the cell invasion; wound healing method was used to detect the cell migration; plate cloning assay was employed to detect the clone formation; flow cytometry was used to detect the cell cycles; Western blotting was used to detect the expressions of A2B5, CD133, nestin, Sox2, Oct-4 and Nanog, as well as phosphatidylinositol kinase (PI3K)-protein kinase B (AKT)-mammalian target of rapamycin (mTOR) signal pathway key proteins (PI3K, AKT, phosphorylated-AKT, and mTOR). Results:The CRNDE mRNA expression in U251s group was significantly higher than that in U251 group ( P<0.05). As compared with U251s group, U251s-CRNDE group had significantly decreased cell viability, cell invasion and cell migration, statistically smaller number of cell colony, significantly decreased proportion of cells in G2/M phase, significantly increased proportion of cells in G1/G0 phase ( P<0.05); and the same trend was noted between U251 group and U251-CRNDE group. As compared with U251s group, U251s-CRNDE group had significantly decreased expressions of CD133, nestin, Sox2, Oct-4, PI3K, AKT, phosphorylated-AKT, and mTOR ( P<0.05). Conclusion:The CRNDE expression is increased in glioma stem cells, and down-regulation of CRNDE expression can inhibit the differentiation, metabolism and proliferation of glioma stem cells; and this effect is related to the regulation of PI3K-AKT-mTOR signaling pathway.
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Objective:To investigate the expression and significance of the angiotensin Ⅱ and its receptors in lipopolysaccharide (LPS)-induced acute lung injury and acute kidney injury in rats.Methods:Forty eight Wistar rats were randomly divided into two groups: control group and endotoxin group (LPS group). LPS was injected through tail vein in LPS group, and the same amount of saline was injected through tail vein in control group.Samples were collected at 2 h, 6 h, 12 h and 24 h, respectively.The histopathology of lung and kidney was observed by HE staining.We detected lung wet/dry weight ratio, serum creatinine, urea nitrogen and Ang Ⅱ concentration in plasma, lung and kidney tissues by radioimmunoassay.Western blot and immunohistochemistry were used to detect the expression changes of AT1R and AT2R in lung and kidney tissue.Results:Compared with the control group, the pathology of lung and kidney tissue in LPS group showed different degrees of damage.The lung wet/dry weight ratio, serum creatinine and urea level in LPS group were significantly increased than that in control group( P<0.05). The Ang Ⅱ content in plasma increased significantly at 2 h and 6 h ( P<0.05), and the expression level of Ang Ⅱ in lung and kidney increased significantly at all time points ( P<0.05). The expression of AT1R in lung and kidney decreased significantly ( P<0.05), while the AT2R protein expression increased significantly ( P<0.05). Additionally the correlation analysis showed that the expression level of Ang Ⅱ and AT2R were positively correlated with lung and renal function, while the expression of AT1R was negatively correlated with lung and renal function. Conclusion:LPS results in the damage of lung and kidney function and the change of renin-angiotensin system.The changes of Ang Ⅱ and angiotension receptors were correlated with lung and kidney injury.Ang Ⅱ and angiotension receptors may be involved in LPS induced lung and kidney injury.
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Objective:To explore the values of Z-scores of fetal heart circumference (HC) and heart area (HA) in prediction of homozygous α-thalassemia.Methods:From February 2014 to March 2019, 233 fetuses of 15 to 23 gestation weeks with risk of homozygous α-thalassemia were examined by prenatal ultrasound in the Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region. HC, HA and cardiothoracic ratio (CTR) were measured. HC and HA were converted into Z-scores, respectively. The ROC curves were established and analyzed based on HC Z-score, HA Z-score or CTR respectively to compare their predicting efficacies for fetal homozygous α-thalassemia. Finally, the sensitivity, specificity, positive predictive value and negative predictive value were obtained by the best cutoff values.Results:①Sixty-five fetuses were diagnosed as homozygous α-thalassemia and classified as α-thalassemia group. One hundred and sixty-eight fetuses were mild and normal and were classified as control group. ②HC Z-score, HA Z-score and CTR in the α-thalassemia group were significantly higher than those in the control group, and the differences between the two groups were statistically significant ( P<0.001). ③The area under ROC curve of HA Z-score was the largest compared with HC Z-score and CTR, and the prediction efficacy was the highest ( Z test=2.144 and 2.517 respectively, P<0.05). ④The best cutoff values were HC Z-score>1.67, HA Z-score>2.06 and CTR>0.53. Sensitivities of predicting homozygous α-thalassemia in 15 to 23 gestation weeks were 92.31%, 100% and 89.23%, respectively. Specificities were 94.05%, 95.83% and 93.45%, respectively. Positive predictive values were 84.43%, 89.00% and 84.05%, respectively. Negative predictive values were 96.91%, 100% and 95.57%, respectively. Conclusions:Fetal heart HC Z-score and HA Z-score are safe and effective novel ultrasonic indexes for predicting homozygous α-thalassemia. Especially compared with traditional CTR, HA Z-score has a significantly higher predicting efficacy, which can improve the detection rate of homozygous α-thalassemia and reduce the need for invasive examination.
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Objective:To construct Z-score ranges for normal fetal middle cerebral artery peak systolic velocity(MCA-PSV).Methods:From May 2017 to October 2019, 865 normal singleton fetuses of 10th to 40th gestational weeks underwent prenatal ultrasound in the Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region. Using fetal biometric parameters as independent variables, and measurement of MCA-PSV on standard section as dependent variables, the regression analyses of the mean(M) and the standard deviation(SD) for each parameter were calculated separately and then the best fitting equation was selected. A group of diseases which might cause the abnormal MCA-PSV were assessed using these standards.Results:①Strong correlations were found between MCA-PSV and fetal biometric parameters ( r=0.935-0.939, P<0.001). ②Quadratic or cubic regression equations were fitted to the models of the means of the MCA-PSV, whereas linear equations were fitted to the SDs. ③In these case groups, intrauterine growth restriction, severe preeclampsia, intrauterine infection and homozygous α-thalassemia-1 demonstrated Z-score>2 reflective of increased MCA-PSV with varying degrees, especially with the homozygous α-thalassemia-1 fetus being the most significant (17/20, 85%). Conclusions:The calculation of Z-score for MCA-PSV as a function of fetal biometric parameters is intuitive and simple, it can be used as an important indicator especially for homozygous α-thalassemia-1.
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Objective To construct a human renal epithelial cell line HEK293T by CRISPR-Cas9-based site-directed knock-in of vascular endothelial growth factor 165 (VEGF165) gene, and avoid the off-target effect caused by lentivirus infection. Methods The VEGF165 expression vector with homologous arm (pUCm-T-VEGF165 plasmid) and the sgRNA expression vector [pSpCas9(BB)-2A-Puro-sgRNA plasmid] were designed and constructed based on the DNA sequence of the EZH2 gene, and then co-transfected into HEK293T cells. The expression of VEGF165 mRNA was detected by qPCR and the expressions of VEGF165 proteins were detected by Western Blot. Results The qPCR and Western Blot results showed that, comparing with the control, the pUCm-T-VEGF165 plasmid and pSpCas9(BB)-2A-Puro-sgRNA plasmid, the expression of the co-transfection plasmid were significantly increased, i.e. 3.42±0.30 vs. 1.02±0.21, 1.13±0.16 and 0.98±0.18 for the VEGF165 mRNA level (all P<0.01), and 1.13±0.16 vs. 1.02±0.06, 0.88±0.03 and 0.80±0.05 for the VEGF165 protein level (all P<0.01), respectively. Besides, the expression of EZH2 was significantly down-regulated, i.e. 0.14±0.06 vs. 1.08±0.11, 1.02±0.12 and 1.13±0.16 for the EZH2 mRNA level (all P<0.01), and 0.23±0.03 vs. 1.05±0.13, 0.91±0.04 and 0.81±0.06 for the EZH2 protein level (all P<0.01), respectively. This result showed that the VEGF165 was successfully inserted into the EZH2 genome, interfering the EZH2 expression. Conclusions VEGF165 gene can be successfully knocked into HEK293T cells by CRISPR/Cas9 system.
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Objective To research the effect of alkylation of glycerol phosphate synthase (AGPS) in isoproterenol (ISO) induced rat cardiac hypertrophy. Methods The pathological cardiac hypertrophy rat model was constructed by ISO intraperitoneal injection. Twelve healthy Sprague-Dawley rats (120~150 g) were divided into ISO group and control group randomly. In the ISO group, rats were injected with ISO (3 mg/kg) per day for two consecutive weeks. In the control group, rats were injected with normal saline (3 mg/kg) per day for two consecutive weeks. Changes of left ventricular diastolic diameter, left ventricular posterior wall thickness, left ventricular ejection fraction, left ventricular short-axis shortening rate and left ventricular mass were detected by echocardiography. The cross-sectional area of myocardial cells in rats was measured by hematoxylin-eosin staining. The expression of hypertrophic factors [atrial natriuretic peptide (ANP), myosin light chain-2V (MLC-2V), α-myosin heavy chain (α-MHC)] and AGPS were detected by Western Blot and real-time quantitative PCR (qPCR). Results The results of echocardiography showed that the cardiac hypertrophy rat model was successfully constructed. The results of hematoxylin-eosin staining showed that the myocardial cross-sectional area in the ISO group was significantly larger than that of the control group. The Western Blot and qPCR results indicated that the relative expression of protein and mRNA of hypertrophic factor and AGPS in the ISO group were both up-regulated comparing with that of the control group, and the differences were statistical significance (all P<0.05). Conclusions The rat model of pathological cardiac hypertrophy with up-regulated AGPS expression was successfully constructed providing a theoretical basis for further study on the role of AGPS in pathogenesis of pathological cardiac hypertrophy.
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The striatum and globus pallidus are principal nuclei of the basal ganglia. Nissl- and acetylcholinesterase-stained sections of the tree shrew brain showed the neuroanatomical features of the caudate nucleus (Cd), internal capsule (ic), putamen (Pu), accumbens, internal globus pallidus, and external globus pallidus. The ic separated the dorsal striatum into the Cd and Pu in the tree shrew, but not in rats and mice. In addition, computer-based 3D images allowed a better understanding of the position and orientation of these structures. These data provided a large-scale atlas of the striatum and globus pallidus in the coronal, sagittal, and horizontal planes, the first detailed distribution of parvalbumin-immunoreactive cells in the tree shrew, and the differences in morphological characteristics and density of parvalbumin-immunoreactive neurons between tree shrew and rat. Our findings support the tree shrew as a potential model for human striatal disorders.
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Animals , Male , Mice , Rats , Acetylcholinesterase , Metabolism , Brain Mapping , Corpus Striatum , Cell Biology , Metabolism , Globus Pallidus , Cell Biology , Metabolism , Imaging, Three-Dimensional , Mice, Inbred C57BL , Models, Neurological , Neurons , Metabolism , Parvalbumins , Metabolism , Rats, Sprague-Dawley , Statistics, Nonparametric , TupaiidaeABSTRACT
Objective To evaluate the efficacy of the ratio of the fetal cardiac diameter to biparietal diameter( CBR) as a predictor of homozygous α-thalassemia-1 . Methods Single mid-pregnancies ( 15-22weeks) at risk of homozygous α-thalassemia-1 were enrolled . A total of 251 singleton pregnancies were recruited ,in which 63 cases were homozygous α-thalassemia-1 fetuses and the rest were unaffected . The CBR and cardiothoracic ratio(CTR) were measured by two-dimensional ultrasound . Then the accuracy of these variables were analyzed and compared with each other by ROC curves . Results ①The CBR and CTR in affected fetuses were significantly higher than those in the unaffected( P <0 .01) . ②With CBR>0 .43 and CTR > 0 .52 as the best cut off values ,the sensitivity and specificity of predicting homozygous α-thalassemia-1 fetuses in 15-22 gestational weeks were 95 .74% , 92 .06% and 94 .15% , 85 .71% , respectively ;the area under ROC curve were compared with Z test and there was no significant difference between them ( Z = 1 .500 , P = 0 .1335) . ③ When CBR and CTR were combined ,the sensitivity and specificity of the prediction were significantly increased ( the sensitivity of series experiment : 99 .75% ,the specificity of parallel experiment : 98 .87% ) . Conclusions CBR is a novel , effective and noninvasive predictor of homozygous α-thalassemia-1 in mid-pregnancy whose prediction efficiency is the same as traditional CTR . The measurement of CBR is easier to standardize and is not affected by thoracic lesions such as pleural cavity ,pericardial effusion and skeletal dysplasia . If combined with CTR ,it may play an important role in improving the prenatal detection rate of homozygous α-thalassemia-1 fetuses .
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Objective To compare the difference in heart rate variability (HRV)between patients with dementia with Lewy bodies(DLB)and those with Parkinson disease dementia(PDD)and to identify the influencing factors.Methods We retrospectively enrolled 30 patients with DLB(DLB group)and 41 patients with PDD(PDD group)at the outpatient and inpatient services of our hospital from January 2010 to December 2017 in this study,and further recruited 119 normal elderly individuals to serve as the control group.HRV was recorded with 24 h dynamic electrocardiogram and compared between the DLB group and the PDD group.Time domain measures including standard deviation of all normal to normal(NN)intervals(SDNN)and square root of the mean squared differences of successive N N intervals(rMSSD)and frequency domain measures including total power(TP),low frequency(LF),and high frequency(HF)were analyzed with a customized program.The levodopa equivalent dose (LED),mini-mental state examination (MMSE),Montreal Cognitive Assessment (MoCA),Hoehn-Yahr stage(H-Y stage)and the unified Parkinson's disease rating scale Ⅲ (UPDRS Ⅲ),and the Alzheimer's disease Cooperative Study-Activities of Daily Living scale(ADCS-ADL)were assessed in DLB and PDD patients to investigate the influencing factors.Results SDNN,TP,and LF in the DLB group and the PDD group were significantly lower than in the control group (F =14.154,4.742,4.897,P<0.05).Compared with the control group,rMSSD decreased in the DLB group,but had no significant difference in the PDD group(P>0.05).The DLB group and the PDD group did not differ in HF from the control group (P > 0.05).There was no significant difference in any HRV indexes between DLB and PDD patients(P >0.05).Correlation analysis showed no correlation of HRV with cognitive level(MMSE,MoCA),motor disturbance degree (UPDRS Ⅲ,H-Yahr stage),daily living ability(ADCS-ADL),or dosage of anti-PD drugs (each P > 0.05).Conclusions DLB and PDD patients present similar impairments in autonomic nervous system function,which are not associated with disease severity.