MAGED4 Expression in Glioma and Upregulation in Glioma Cell Lines with 5-Aza-2'-Deoxycytidine Treatment

Zhang, Qing-Mei,Shen, Ning,Xie, Sha,Bi, Shui-Qing,Luo, Bin,Lin, Yong-Da,Fu, Jun,Zhou, Su-Fang,Luo, Guo-Rong,Xie, Xiao-Xun,Xiao, Shao-Wen Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.8

Melanoma-associated antigen (MAGE) family genes have been considered as potentially promising targets for anticancer immunotherapy. MAGED4 was originally identified as a glioma-specific antigen. Current knowledge about MAGED4 expression in glioma is only based on mRNA analysis and MAGED4 protein expression has not been elucidated. In the present study, we investigated this point and found that MAGED4 mRNA and protein were absent or very lowly expressed in various normal tissues and glioma cell line SHG44, but overexpressed in glioma cell lines A172,U251,U87-MG as well as glioma tissues, with significant heterogeneity. Furthermore, MAGED4 protein expression was positively correlated with the glioma type and grade. We also found that the expression of MAGED4 inversely correlated with the overall methylation status of the MAGED4 promoter CpG island. Furthermore, when SHG44 and A172 with higher methylation were treated with the DNA demethylating agent 5-aza-2'-deoxycytidine (5-AZA-CdR) reactivation of MAGED4 mRNA was mediated by significant demethylation in SHG44 instead of A172. However, 5-AZA-CdR treatment had no effect on MAGED4 protein in both SHG44 and A172 cells. In conclusion, MAGED4 is frequently and highly expressed in glioma and is partly regulated by DNA methylation. The results suggest that MAGED4 might be a promising target for glioma immunotherapy combined with 5-AZA-CdR to enhance its expression and eliminate intratumor heterogeneity.

Aqueous two-phase extraction to separate alpha-ketoglutaric acid from fermentation broth by combining crystallization

Xueqi Shi,Weiyi Su,Hongyu Zhang,Jing Fang,Ning Xu,Yanjun Jiang,Hao Li 한국공업화학회 2022 Journal of Industrial and Engineering Chemistry Vol.115 No.-

Alpha-ketoglutaric acid (a-KG) is a crucial intermediate with broad application prospects. However, thedownstream separation of a-KG from fermentation broth with low concentration and complex componentshas become the bottleneck of large-scale production. In this study, a-KG was separated from theactual fermentation broth by aqueous two-phase system (ATPS) and crystallization. The optimal ATPScomposed of acetone/(NH4)2SO4 was selected by studying the phase diagram and a-KG distributionbehavior in different organic solvents and salts/sugars. Under the optimized conditions, the purity ofa-KG products from crude fermentation broth reached 98.08%, and the one-pass extraction efficiencywas 90.52%. The removal of cell debris and soluble proteins increased with the increase of acetone and(NH4)2SO4 concentrations, with the removal ratios reaching up to 99.44% and 86.54%. In addition,92.56% of the glucose was distributed in the aqueous phase. Although the a-KG fermentation broth ismore complex than the whole-cell biotransformation, impurities in the crude fermentation broth couldbe efficiently removed and a higher a-KG distribution coefficient was obtained. Meanwhile, the recovered(NH4)2SO4 could be reused for fermentation medium. This strategy provides an economical and effectivemethod combined with upstream biosynthesis to recover a-KG and other bioproducts from crude fermentationbroth.

An update of preimplantation genetic diagnosis in gene diseases, chromosomal translocation, and aneuploidy screening

Chang, Li-Jung,Chen, Shee-Uan,Tsai, Yi-Yi,Hung, Chia-Cheng,Fang, Mei-Ya,Su, Yi-Ning,Yang, Yu-Shih The Korean Society for Reproductive Medicine 2011 Clinical and Experimental Reproductive Medicine Vol.38 No.3

Preimplantation genetic diagnosis (PGD) is gradually widely used in prevention of gene diseases and chromosomal abnormalities. Much improvement has been achieved in biopsy technique and molecular diagnosis. Blastocyst biopsy can increase diagnostic accuracy and reduce allele dropout. It is cost-effective and currently plays an important role. Whole genome amplification permits subsequent individual detection of multiple gene loci and screening all 23 pairs of chromosomes. For PGD of chromosomal translocation, fluorescence $in-situ$ hybridization (FISH) is traditionally used, but with technical difficulty. Array comparative genomic hybridization (CGH) can detect translocation and 23 pairs of chromosomes that may replace FISH. Single nucleotide polymorphisms array with haplotyping can further distinguish between normal chromosomes and balanced translocation. PGD may shorten time to conceive and reduce miscarriage for patients with chromosomal translocation. PGD has a potential value for mitochondrial diseases. Preimplantation genetic haplotyping has been applied for unknown mutation sites of single gene disease. Preimplantation genetic screening (PGS) using limited FISH probes in the cleavage-stage embryo did not increase live birth rates for patients with advanced maternal age, unexplained recurrent abortions, and repeated implantation failure. Polar body and blastocyst biopsy may circumvent the problem of mosaicism. PGS using blastocyst biopsy and array CGH is encouraging and merit further studies. Cryopreservation of biopsied blastocysts instead of fresh transfer permits sufficient time for transportation and genetic analysis. Cryopreservation of embryos may avoid ovarian hyperstimulation syndrome and possible suboptimal endometrium.

Effect of wind time on the dispersal capacity of Trichogramma dendrolimi Matsumura (Hymenoptera Trichogrammatidae)

Jin-cheng Zhou,Qian-jin Dong,Tong-shu Zhang,Li-jia Duan,Su-fang Ning,Quanquan Liu,Yuan-yuan Li,Chun-xue Li,Hui Dong 한국응용곤충학회 2019 Journal of Asia-Pacific Entomology Vol.22 No.3

The efficacy of Trichogramma against pests basically depends on the dispersal capacity of the wasps. A releaserecapture method was used in this study to evaluate the effect of relative wind speed (RWS) during the nighttime and the daytime on the dispersal capacity of postrelease population of Trichogramma dendrolimi Matsumura. Two repeated measurements were carried out, using sentinel cards with the host eggs and yellow sticky card traps on the third and the sixth day after wasp release. The results showed that: (1) both the number of recaptured wasps and parasitized eggs increased with the RWS in the daytime during the first three days. The number of parasitized eggs also increased with the RWS in the daytime over the latter three days. The distances that encompassed 98% recaptured T. dendrolimi adults (X 98 ) at the first three days was quadratically increased by the RWS of the daytime. (2) The number of wasps recaptured decreased with the distance from the releasing point during the first three days, but it was not influenced by the distance from the releasing point during the latter three days. (3) The average dispersal radius of the wasps during the first three days was shorter than during the latter three days. The present results will be helpful to improve release techniques based on the wind effects at different times.

Selection and evaluation of RT-qPCR reference genes for expression analysis in the tiny egg parasitoid wasp, Trichogramma dendrolimi matsumura (Hymenoptera: Trichogrammatidae)

Huo Liang-xiao,Bai Xue-ping,Che Wu-nan,Ning Su-fang,Lv Lin,Zhang Lisheng,Zhou Jin-cheng,Dong Hui 한국응용곤충학회 2022 Journal of Asia-Pacific Entomology Vol.25 No.2

The egg parasitoid, Trichogramma spp., is an important biological control agent used against a broad range of Lepidopteran pests in agriculture and forestry. The biology of Trichogramma has been studied in details. Further studies should focus on the molecular mechanisms of Trichogramma by qualifying the expression of related genes It is critical to select appropriate reference genes for normalizing RT-qPCR results and establishing a robust method for quantifying target gene expression. This study aimed to identify and validate appropriate reference genes for use in RT-qPCR analysis of Trichogramma dendrolimi. Ten candidate housekeeping genes, namely betaactin (ACTIN), forkhead box O (FOXO), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), heat shock pro tein 90 (HSP90), ribosomal protein L10a (RPL10a), L18 (RPL18), L28 (RPL28), S13 (RPS13), S15 (RPS15), and superoxide dismutase (SOD), were tested for their suitability as reference genes for developmental stage (3rd, 4th, 5th, 6th, 7th, 8th, 9th, and 10th day after parasitization), tissue (head, thorax, and abdomen of adults), sex of adults (male and female), and temperature (17℃, 25℃, and 32℃). According to the GeNorm analysis, a robust analysis should involve using an appropriate combination of reference genes, namely, at least three genes for different development stages, two genes for different tissues, two genes for different sex, and two genes for different temperatures, respectively. According to the RelFinder method by the integrated results of GeNorm, NormFinder, BestKeeper, and the ΔCt method, we identified the developmental stage-specific reference genes SOD, GAPDH, and ACTIN; tissue-specific reference genes RPL18 and RPS15; sex-specific reference genes RPL18 and SOD; and temperature-specific reference genes RPL18 and RPL10a. This study provides a standardized procedure for the quantification of gene expression in T. dendrolimi and will be helpful for future biological control programs using Trichogramma wasps.

Predictive Value of Exercise Blood Pressure Changes for Orthostatic Hypotension in Patients With Parkinson’s Disease

Yi Qin,Zhao-hui Jin,Zhen-ying Zhang,Ke-ke Chen,Xin Yu,Hong-jiao Yan,Rui-dan Wang,Yuan Su,Ai-xian Liu,Jia-ning Xi,Bo-yan Fang 대한신경과학회 2023 Journal of Clinical Neurology Vol.19 No.1

Background and Purpose Orthostatic hypotension (OH) is common in patients with Parkinson’s disease (PD). Early recognition OH is required with sensitive assessments. The purpose of this study was to determine whether blood pressure (BP) changes during exercise can predict the occurrence of OH in PD. Methods This prospective cohort study included 80 consecutive patients with PD. All patients agreed to participate in a baseline evaluation and cardiopulmonary exercise test (CPET). According to the initial active standing test (AST), those without OH (PD-nonOH) at baseline had their AST results followed up for 6 months. The main outcome was defined as whether patients without OH at baseline would develop OH after 6 months. Logistic regression analysis was applied to identify the relevant variables. A nomogram was constructed based on clinical features and identified variables. The concordance index (C-index) and area under the receiver operating characteristic curve (AUC) were used to evaluate the accuracy and predictive ability of the nomogram, respectively. Results CPET results indicated that peak load, peak heart rate, heart rate recovery at 1 min, and systolic BP change (ΔSBP) were lower in those with OH than in the PD-nonOH group (p<0.05) at baseline. Logistic regression analysis indicated that peak load and ΔSBP during CPET had significant effects on OH (p<0.05). Age, sex, peak load, and ΔSBP were used to construct the nomogram model (C-index=0.761). The prediction model had an AUC of 0.782 (95% confidence interval=0.649–0.889) and a specificity and sensitivity of 70.0% and 81.8%, respectively. Conclusions This study has identified predictive factors for OH development in patients with PD. CPET could be used as a complementary examination to identify patients at a high risk of OH.