Cloning and Characterization of a Na+/H+ Antiporter Gene of the Moderately Halophilic Bacterium Halobacillus aidingensis AD-6T

Ya Jie Zou,Li Fu Yang,Lei Wang,Su Sheng Yang 한국미생물학회 2008 The journal of microbiology Vol.46 No.4

A gene encoding a Na+/H+ antiporter was obtained from the genome of Halobacillus aidingensis AD-6T, which was sequenced and designated as nhaH. The deduced amino acid sequence of the gene was 91% identical to the NhaH of H. dabanensis, and shared 54% identity with the NhaG of Bacillus subtilis. The cloned gene enable the Escherichia coli KNabc cell, which lack all of the major Na+/H+ antiporters, to grow in medium containing 0.2 M NaCl or 10 mM LiCl. The nhaH gene was predicted to encode a 43.5 kDa protein (403 amino acid residues) with 11 putative transmembrane regions. Everted membrane vesicles prepared from E. coli KNabc cells carrying NhaH exhibited Na+/H+ as well as Li+/H+ antiporter activity, which was pH-dependent with the highest activity at pH 8.0, and no K+ /H+ antiporter activity was detected. The deletion of hydrophilic C-terminal amino acid residues showed that the short C-terminal tail was vital for Na+/H+ antiporter activity.

Effects of Supplementing Different Combinations of Nitrogen Supplements on Digestibility and Performance in Sheep Fed Wheat Straw Diets

Li, Yong,Li, Defa,Xing, Jianjun,Li, Shengli,Han, Yang,Han, In K. Asian Australasian Association of Animal Productio 2002 Animal Bioscience Vol.15 No.10

Thirty-six (20 wethers and 16 rams) adult Gansu highland finewool sheep with an average liveweight of $19.42{\pm}1.83kg$ were used. The sheep were allocated to 4 treatments and confined in individual feeding stalls. The 80 day feeding trial was divided into 2 periods of 38 and 42 days. The sheep had access to clean water and were fed chopped wheat straw ad libitum. Daily supplemented diets of 150 g for the first period and 200 g for the later period were provided. The 4 types of supplements were: Wheat bran type (I), which is the traditional feeding practice in the area and the control group; Linseed cake type (II); Linseed cake+poultry litter type (III); Poultry litter+distiller grains type (IV). The digestibility trial was conducted using a total collection method at the end of the first period in the feeding trial. The results showed that the digestibilities of DM, OM, CP, CF, NDF and ADF in group II and III were increased to more than 50%. CP and NDF digestibility in group II were significantly increased by 23.6% and 25.5% respectively over group I (p<0.01) and by 10.1% and 13.1% respectively over group III (p<0.05). The digestibility of DM, OM, CP, CF, NDF and ADF in group IV was lower than group I by 4.4 to 8.4% (p>0.05). Compared to group I, group II and III increased straw intake by 17.6% (p<0.01) and 10.8% (p<0.05). The ADG increased from 16.2 g in group I to 45.3 g and 32.8 g (p<0.01) in group II and III respectively. The ratio of supplement intake to gain decreased from 10.7 in group I to 3.9 and 5.4 (p<0.01) in group II and III respectively. The ratio of supplement cost to gain also decreased by 60.0% and 64.3%, respectively. The wool growth rate and wool strength in these 2 groups were noticeably improved. This significantly enhanced the wool's textile value. The above parameters in group IV were lower than in group I, except the ratio of supplement cost to gain. This research indicates that local low quality nitrogen resources could be used effectively and economically to improve the utilization of wheat straw fed to sheep on smallholder farms under harsh loess plateau conditions in the winter.

Advanced glycation end products promote meniscal calcification by activating the mTOR-ATF4 positive feedback loop

Yang Sheng,Xie JiaJun,Pan ZhiJie,Guan HongMei,Tu YueSheng,Ye YuanJian,Huang ShouBin,Fu ShiQiang,Li KangXian,Huang ZhiWei,Li XiaoQi,Shi ZhanJun,Li Le,Zhang Yang 생화학분자생물학회 2024 Experimental and molecular medicine Vol.56 No.-

The meniscus is vital for maintaining knee homeostasis and function. Meniscal calcification is one of the earliest radiological indicators of knee osteoarthritis (KOA), and meniscal calcification is associated with alterations in biomechanical properties. Meniscal calcification originates from a biochemical process similar to vascular calcification. Advanced glycation end products (AGEs) and their receptors (RAGEs) reportedly play critical roles in vascular calcification. Herein, we investigated whether targeting AGE-RAGE is a potential treatment for meniscal calcification. In our study, we demonstrated that AGE-RAGE promotes the osteogenesis of meniscal cells and exacerbates meniscal calcification. Mechanistically, AGE-RAGE activates mTOR and simultaneously promotes ATF4 accumulation, thereby facilitating the ATF4-mTOR positive feedback loop that enhances the osteogenic capacity of meniscal cells. In this regard, mTOR inhibits ATF4 degradation by reducing its ubiquitination, while ATF4 activates mTOR by increasing arginine uptake. Our findings substantiate the unique role of AGE-RAGE in the meniscus and reveal the role of the ATF4-mTOR positive feedback loop during the osteogenesis of meniscal cells; these results provide potential therapeutic targets for KOA.

Survival Benefit of Tamoxifen in Estrogen Receptor-Negative and Progesterone Receptor-Positive Low Grade Breast Cancer Patients

Li-Heng Yang,Hsin-Shun Tseng,Che Lin,Li-Sheng Chen,Shou-Tung Chen,Shou-Jen Kuo,Dar-Ren Chen 한국유방암학회 2012 Journal of breast cancer Vol.15 No.3

Purpose: This study aimed to analyze the efficacy and prognostic significance of adjuvant tamoxifen in breast cancer patients with various hormone receptor statuses. Methods: Typically, 1,260 female breast cancer patients were recruited in this study. The correlation between estrogen receptor (ER)/progesterone receptor (PR) phenotypes and clinical characteristics was investigated, and the survival rate was assessed after 5-year follow-up. Results: The 5-year overall survival (85%) was better in women under the age of 50 years. Patients with ER+/PR+ tumors had a better 5-year survival rate (94%); those with ER-/PR- tumors experienced the worst outcome (74% survival rate); whereas singlepositive cases were in between. In 97 out of 128 patients with ER-/PR+ tumors, tamoxifen was given as adjuvant hormonal therapy, and it increased the survival benefit in the lower grade group in terms of overall survival and disease-free survival (p=0.01 and p=0.03, respectively). Conclusion: For high-grade tumors with ER-/PR+, adjuvant tamoxifen therapy may have no survival benefit, whereas for the patients with low-grade ER-/PR+ tumors, adjuvant tamoxifen therapy is highly suggestive.

Expression profiles of circular RNAs in sheep skeletal muscle

Cao, Yang,You, Shuang,Yao, Yang,Liu, Zhi-Jin,Hazi, Wureli,Li, Cun-Yuan,Zhang, Xiang-Yu,Hou, Xiao-Xu,Wei, Jun-Chang,Li, Xiao-Yue,Wang, Da-Wei,Chen, Chuang-Fu,Zhang, Yun-Feng,Ni, Wei,Hu, Sheng-Wei Asian Australasian Association of Animal Productio 2018 Animal Bioscience Vol.31 No.10

Objective: Circular RNAs (circRNAs) are a newfound class of non-coding RNA in animals and plants. Recent studies have revealed that circRNAs play important roles in cell proliferation, differentiation, autophagy and apoptosis during development. However, there are few reports about muscle development-related circRNAs in livestock. Methods: RNA sequencing analysis was employed to identify and annotate circRNAs from longissimus dorsi of sheep. Reverse transcription followed by real-time quantitative (q) polymerase chain reaction (PCR) analysis verified the presence of these circRNAs. Targetscan7.0 and miRanda were used to analyse the interaction of circRNA-microRNA (miRNA). To investigate the function of circRNAs, an experiment was conducted to perform enrichment analysis hosting genes of circRNAs using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. Results: About 75.5 million sequences were obtained from RNA libraries of sheep skeletal muscle. These sequences were mapped to 729 genes in the sheep reference genome. We identified 886 circRNAs, including numerous circular intronic RNAs and exonic circRNAs. Reverse transcription PCR (RT-PCR) and DNA sequencing analysis confirmed the presence of several circRNAs. Real-Time RT-PCR analysis exhibited resistance of sheep circRNAs to RNase R digestion. We found that many circRNAs interacted with muscle-specific miRNAs involved in growth and development of muscle, especially circ776. The GO and KEGG enrichment analysis showed that hosting genes of circRNAs was involved in muscle cell development and signaling pathway. Conclusion: The study provides comprehensive expression profiles of circRNAs in sheep skeletal muscle. Our study offers a large number of circRNAs to facilitate a better understanding of their roles in muscle growth. Meanwhile, we suggested that circ776 could be analyzed in future study.

Evaluation of Several Screening Approaches for Detection of Cervical Lesions in Rural Shandong, China

Zong, Li-Ju,Zhang, You-Zhong,Yang, Xing-sheng,Jiang, Jie,Cui, Bao-Xia,Qiao, Yun-Bo,Li, Li,Jiang, Kan,Zhang, Wen-Jing,Kong, Bei-Hua,Shen, Keng Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.5

Purpose: The study was designed to: (1) investigate the prevalence of high-risk human papillomavirus (HR-HPV) infection and cervical neoplasia; and (2) evaluate clinical performance of visual inspection with acetic acid/ Lugol's iodine (VIA /VILI), Pap smear, high-risk human papillomavirus (HR-HPV) DNA test for detecting cervical intraepithelial neoplasia grade 2 or worse (CIN2+) and (3) explore appropriate screening approach in rural areas of Shandong Province. Materials and Methods: A total of 3,763 eligible women from Yiyuan County in Yimeng mountainous areas of rural Shandong, China, were enrolled and underwent Pap smear, HR-HPV DNA testing by Hybrid Capture 2 (HC2), and VIA /VILI tests. Women positive in any test were referred to colposcopy and biopsy as indicated. Results: The prevalence of HR-HPV infection among all enrolled women was 11.1% and that in healthy women was 9.9%. In total 33 cases of CIN1, 16 cases of CIN2, 6 cases of CIN3 but none of cervical cancer were detected and the crude prevalence of CIN2+ was 0.58%. For detecting CIN2+, the sensitivity of HR-HPV DNA testing, VIA/VILI, Pap smear was 90.9%, 77.3%, 81.8%, respectively. Pap smear had the best specificity of 98.2%, followed by HR-HPV DNA testing with specificity of 89.4%, VIA/VILI had the lowest specificity of 81.2%. Colposcopy referral rate of HR-HPV DNA testing, VIA/VILI, Pap smear was 11.1%, 18.5%, 2.3%, respectively. Conclusions: Our results suggest that HR-HPV DNA testing alone might be appropriate for primary cervical cancer screening in rural low-resource areas of Shandong Province, China.

Time-related Mapping of Quantitative Trait Loci Controlling Dry Matter Accumulation and Partitioning in Rice

Shao Bo Li,Zhi Hong Zhang,Shao Qing Li,Yang Sheng Li,You Lin Zhu,Ying Guo Zhu 한국유전학회 2008 Genes & Genomics Vol.30 No.4

Grain yield is a result of dry matter accumulation and partitioning in rice (Oryza sativa L.). To characterize the genetic basis of dynamics of traits representing dry matter accumulation and partitioning as well as their relationships with each other, composite interval mapping of quantitative trait loci (QTL) was conducted based on time-related phenotypic data obtained from a population of 258 recombinant inbred lines (F12), which was derived from a cross between Lemont (japonica var.) and Teqing (indica var.). A total of twenty-three QTL were identified for leaf weight, stem weight and panicle weight at four measuring stages after heading. However, these three dynamic traits were observed to be controlled by different QTL (genes) at different growth stages. An extreme example was that at least three types of QTL (genes) might be involved in the development of panicle weight. Growth duration, which is closely related to dry matter accumulation and partitioning, was also identified to be significantly affected by four QTL. The close correlations among the studied four traits were explained to some extent by six co-located QTL. Nevertheless, it would be of great importance to distinguish the pleiotropic or linked effects of the co-located QTL. In addition, the chromosome regions or QTL identified in this study were compared with those previously reported for the same or related traits with the same RIL population.

Gene Silencing of β-catenin by RNAi Inhibits Proliferation of Human Esophageal Cancer Cells by Inducing G0/G1 Cell Cycle Arrest

Wang, Jin-Sheng,Ji, Ai-Fang,Wan, Hong-Jun,Lu, Ya-Li,Yang, Jian-Zhou,Ma, Li-Li,Wang, Yong-Jin,Wei, Wu Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.6

Objectives: The aim of the present study was to explore mechanisms underlying the effects of down-regulating ${\beta}$-catenin expression on esophageal carcinoma (EC) cells. Methods: Cell cycle distribution and apoptosis were determined using flow cytometry and annexin V apoptosis assay, respectively. Transmission electron microscopy (TEM) was used to examine changes in ultrastructure, while expression of cyclin D1 protein and mRNA was detected by western blot and real-time PCR. Proliferating cell nuclear antigen (PCNA) and extracellular signal-regulated kinase (ERK) 1-2 were evaluated by Western blot analysis. PCNA labeling index (LI) was determined by immunocytochemistry. Results: Compared with pGen-3-con transfected and Eca-109 cells, the percentage of G0/G1-phase pGen-3-CTNNB1 transfected cells was obviously increased (P<0.05), with no significant difference among the three groups with regard to apoptosis (P>0.05). pGen-3-CTNNB1 transfected cells exhibited obvious decrease in cyclin D1 mRNA and protein expression (P<0.05) and the ultrastructure of Eca-109 cells underwent a significant change after being transfected with pGen-3-CTNNB1, suggesting that down-regulating ${\beta}$-catenin expression can promote the differentiation and maturation. The expression of PCNA and the ERKI/2 phosphorylation state were also down-regulated in pGen-3-CTNNB1 transfected cells (P<0.05). At the same time, the PCNA labeling index was decreased accordingly (P<0.05). Conclusion: Inhibition of EC Eca-109 cellproliferation by down-regulating ${\beta}$-catenin expression could improve cell ultrastructure by mediating blockade in G0/G1 through inhibiting cyclin D1, PCNA and the MAPK pathway (p-ERK1/2).

Serum Peroxiredoxin3 is a Useful Biomarker for Early Diagnosis and Assessemnt of Prognosis of Hepatocellular Carcinoma in Chinese Patients

Shi, Liang,Wu, Li-Li,Yang, Jian-Rong,Chen, Xiao-Fei,Zhang, Yi,Chen, Zeng-Qiang,Liu, Cun-Li,Chi, Sheng-Ying,Zheng, Jia-Ying,Huang, Hai-Xia,Yu, Fu-Jun,Lin, Xiang-Yang Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.7

Background: Recently, peroxiredoxin3 (PRDX3) was identified as a novel molecular marker for the progression of hepatocellular carcinoma (HCC). However, its potential clinical application as a serum marker for the early diagnosis and prognosis of HCC has not been investigated. Methods: PRDX3, alpha-fetaprotein (AFP), and other biochemical parameters were measured in serum samples from 297 Chinese patients, including 96 with HCC, 98 with liver cirrhosis (LC), and 103 healthy controls (HCs). Correlations between serum PRDX3 expression and clinicopathological variables and the relationship between serum PRDX3 expression and prognosis were analyzed. Results: Serum PRDX3 was significantly higher in HCC patients than in the LC and HC groups. The sensitivity and specificity of serum PRDX3 for the diagnosis of HCC were 85.9% and 75.3%, respectively, at a cutoff of 153.26 ng/mL, and the area under the curve was 0.865. Moreover, serum PRDX3 expression was strongly associated with AFP level, tumor diameter, TNM stage, and portal vein invasion. Kaplan-Meier curve analysis revealed that HCC patients with high serum PRDX3 expression had a shorter median survival time than those with low PRDX3 expression. Moreover, serum PRDX3 expression was an independent risk factor for overall survival. The inverse correlation between serum PRDX3 and patient survival remained significant in patients with early-stage HCC and in those with normal serum AFP levels. Conclusions: Serum PRDX3 can be used as a noninvasive biomarker for the diagnosis and/or prognosis of HCC.

miR-340 Reverses Cisplatin Resistance of Hepatocellular Carcinoma Cell Lines by Targeting Nrf2-dependent Antioxidant Pathway

Shi, Liang,Chen, Zhan-Guo,Wu, Li-li,Zheng, Jian-Jian,Yang, Jian-Rong,Chen, Xiao-Fei,Chen, Zeng-Qiang,Liu, Cun-Li,Chi, Sheng-Ying,Zheng, Jia-Ying,Huang, Hai-Xia,Lin, Xiang-Yang,Zheng, Fang Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.23

Many chemotherapeutic agents have been successfully used to treat hepatocellular carcinoma (HCC); however, the development of chemoresistance in liver cancer cells usually results in a relapse and worsening of prognosis. It has been demonstrated that DNA methylation and histone modification play crucial roles in chemotherapy resistance. Currently, extensive research has shown that there is another potential mechanism of gene expression control, which is mediated through the function of short noncoding RNAs, especially for microRNAs (miRNAs), but little is known about their roles in cancer cell drug resistance. In present study, by taking advantage of miRNA effects on the resistance of human hepatocellular carcinoma cells line to cisplatin, it has been demonstrated that miR-340 were significantly downregulated whereas Nrf2 was upregulated in HepG2/CDDP (cisplatin) cells, compared with parental HepG2 cells. Bioinformatics analysis and luciferase assays of Nrf2-3'-untranslated region-based reporter constructor indicated that Nrf2 was the direct target gene of miR-340, miR-340 mimics suppressing Nrf2-dependent antioxidant pathway and enhancing the sensitivity of HepG2/CDDP cells to cisplatin. Interestingly, transfection with miR-340 mimics combined with miR-340 inhibitors reactivated the Nrf2 related pathway and restored the resistance of HepG2/CDDP cells to CDDP. Collectively, the results first suggested that lower expression of miR-340 is involved in the development of CDDP resistance in hepatocellular carcinoma cell line, at least partly due to regulating Nrf2-dependent antioxidant pathway.