Transmembrane Helix of Novel Oncogene with Kinase-Domain (NOK) Influences Its Oligomerization and Limits the Activation of RAS/MAPK Signaling

Ying-Hua Li,Yin-Yin Wang,Shan Zhong,Zhi-Li Rong,Yong-Ming Ren,Zhi-Yong Li,Shu-Ping Zhang,Zhi-Jie Chang,Li Liu 한국분자세포생물학회 2009 Molecules and cells Vol.27 No.1

Ligand-dependent or independent oligomerization of receptor protein tyrosine kinase (RPTK) is often an essential step for receptor activation and intracellular signaling. The novel oncogene with kinase-domain (NOK) is a unique RPTK that almost completely lacks an ectodomain, expresses intracellularly and activates constitutively. However, it is unknown whether NOK can form oligomer or what function oligomerization would have. In this study, two NOK deletion mutants were generated by either removing the ectodomain (NOKECD) or including the endodomain (NOK-ICD). Co-immunoprecipitation demonstrated that the transmembrane (TM) domain of NOK was essential for its intermolecular interaction. The results further showed that NOK aggregated more closely as lower order oligomers (the dimer- and trimer-sized) than either deletion mutant did since NOK could be cross-linked by both Sulfo-EGS and formaldehyde, whereas either deletion mutant was only sensitive to Sulfo-EGS. Removing the NOK TM domain (NOK-ICD) not only markedly promoted higher order oligomerization, but also altered the subcellular localization of NOK and dramatically elevated the NOK-mediated constitutive activation of extracellular signal-regulated kinase (ERK). Moreover, NOK-ICD but not NOK or NOKECD was co-localized with the upstream signaling molecule RAS on cell membrane. Thus, TM-mediated intermolecular contacting may be mainly responsible for the constitutive activation of NOK and contribute to the autoinhibitory effect on RAS/MAPK signaling.

Iron Determination in Rat Plasma Samples by Inductively Coupled Plasma Emission Spectrometry and Application to Pharmacokinetic Studies

Li, Tie-Fu,Deng, Ying-Jie,Ma, Guang-Li,Jin, Jie,Li, Song Korean Chemical Society 2003 Bulletin of the Korean Chemical Society Vol.24 No.11

This paper describes an inductively coupled plasma emission spectrometric method for the analysis of Fe in rat plasma. Calibration curves were obtained in the range of 0.125-1.50 ${\mu}g{\cdot}mL^{-1}$. The relative standard deviation ranges from 5.93% to 6.80%, and accuracy was between 87.6 and 102.0%. Dilution with water had no influence on the performance of the method, which could then be used to quantify Fe concentration in plasma up to 0.50 ${\mu}g{\cdot}mL^{-1}$. The limit of quantification was 0.10 ${\mu}g{\cdot}mL^{-1}$. At this level, the average relative standard deviation was 6.8%. The results indicate that the method meets the accuracy and precision requirements for the pharmacokinetic studies. The Fe concentration in rat plasma was measured and the main pharmacokinetic parameters were calculated by Topfit 2.0 (GmbH. Shering AG, Godecke AG, Germany).

Ginsenoside Rg3 in combination with artesunate overcomes sorafenib resistance in hepatoma cell and mouse models

Ying-Jie Chen,Jia-Ying Wu,Yu-Yi Deng,Ying Wu,Xiao-Qi Wang,Amy Sze-man Li,Lut Yi Wong,Xiuqiong Fu,Zhi-Ling Yu,Chun Liang 고려인삼학회 2022 Journal of Ginseng Research Vol.46 No.3

Sorafenib is effective in treating hepatoma, but most patients develop resistance to it. STAT3signaling has been implicated in sorafenib resistance. Artesunate (ART) and 20(R)-ginsenoside Rg3 (Rg3)have anti-hepatoma effects and can inhibit STAT3 signaling in cancer cells. This study aimed to evaluatethe effects of Rg3 in combination with ART (Rg3-plus-ART) in overcoming sorafenib resistance, and toexamine the involvement of STAT3 signaling in these effects. Methods: Sorafenib-resistant HepG2 cells (HepG2-SR) were used to evaluate the in vitro anti-hepatomaeffects of Rg3-plus-ART. A HepG2-SR hepatoma-bearing BALB/c-nu/nu mouse model was used to assessthe in vivo anti-hepatoma effects of Rg3-plus-ART. CCK-8 assays and Annexin V-FITC/PI double stainingwere used to examine cell proliferation and apoptosis, respectively. Immunoblotting was employed toexamine protein levels. ROS generation was examined by measuring DCF-DA fluorescence. Results: Rg3-plus-ART synergistically reduced viability of, and evoked apoptosis in HepG2-SR cells, andsuppressed HepG2-SR tumor growth in mice. Mechanistic studies revealed that Rg3-plus-ART inhibitedactivation/phosphorylation of Src and STAT3 in HepG2-SR cultures and tumors. The combination alsodecreased the STAT3 nuclear level and induced ROS production in HepG2-SR cultures. Furthermore, overactivation of STAT3 or removal of ROS diminished the anti-proliferative effects of Rg3-plus-ART, andremoval of ROS diminished Rg3-plus-ART's inhibitory effects on STAT3 activation in HepG2-SR cells. Conclusions: Rg3-plus-ART overcomes sorafenib resistance in experimental models, and inhibition of Src/STAT3 signaling and modulation of ROS/STAT3 signaling contribute to the underlying mechanisms. Thisstudy provides a pharmacological basis for developing Rg3-plus-ART into a novel modality for treatingsorafenib-resistant hepatoma.

The Role of Intestinal Microbiota and Mast Cell in a Rat Model of Visceral Hypersensitivity

( Ying-jie Li ),( Jing Li ),( Cong Dai ) 대한소화기기능성질환·운동학회(구 대한소화관운동학회) 2020 Journal of Neurogastroenterology and Motility (JNM Vol.26 No.4

Background/Aims To explore the role of intestinal flora and mast cells in visceral hypersensitivity (VH). Methods The experimental animals were divided into 4 groups: control group, VH group, VH + VSL#3 group, and VH + ketotifen group. Stool samples were collected from each group (n = 3) for a further analysis using 16S ribosomal DNA gene sequence. Visceral sensitivity was evaluated by abdominal withdrawal reflex (AWR) score. Colon tissues of rats were obtained from each group. Mast cells were detected by toluidine blue staining. The degranulation of mast cells was assessed by transmission electron microscopy. Results VH rat model could successfully be induced by acetic acid enema combined with partial limb restraint method. Compared with rats in the control group, AWR score, number of mast cells, and degranulation of mast cells were increased in the VH rats, which could be reduced by administration of ketotifen or probiotic VSL#3. Clostridium sensu stricto 1 abundance was higher in the VH group compared to the control group, which could be restored by application of probiotic VSL#3. Conclusions Probiotic VSL#3 decreases visceral sensitivity in VH rats. The mechanism may be related to mast cell and intestinal flora. Change of Clostridium sensu stricto 1 abundance may be a basis for VH observed in irritable bowel syndrome and may be prevented by specific probiotic administration. (J Neurogastroenterol Motil 2020;26:529-538)

Mini-Array of Multiple Tumor-associated Antigens (TAAs) in the Immunodiagnosis of Esophageal Cancer

Qin, Jie-Jie,Wang, Xiao-Rui,Wang, Peng,Ren, Peng-Fei,Shi, Jian-Xiang,Zhang, Hong-Fei,Xia, Jun-Fen,Wang, Kai-Juan,Song, Chun-Hua,Dai, Li-Ping,Zhang, Jian-Ying Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

Sera of cancer patients may contain antibodies that react with a unique group of autologous cellular antigens called tumor-associated antigens (TAAs). The present study aimed to determine whether a mini-array of multiple TAAs would enhance antibody detection and be a useful approach in esophageal cancer detection and diagnosis. Our mini-array of multiple TAAs consisted of eleven antigens, p53, pl6, Impl, CyclinB1, C-myc, RalA, p62, Survivin, Koc, CyclinD1 and CyclinE full-length recombinant proteins. Enzyme-linked immunosorbent assays (ELISA) were used to detect autoantibodies against eleven selected TAAs in 174 sera from patients with esophageal cancer, as well as 242 sera from normal individuals. In addition, positive results of ELISA were confirmed by Western blotting. In a parallel screening trial, with the successive addition of antigen to a final total of eleven TAAs, there was a stepwise increase in positive antibody reactions. The eleven TAAs were the best parallel combination, and the sensitivity and specificity in diagnosing esophageal cancer was 75.3% and 81.0%, respectively. The positive and negative predictive values were 74.0% and 82.0%, respectively, indicating that the parallel assay of eleven TAAs raised the diagnostic precision significantly. In addition, the levels of antibodies to seven antigens, comprising p53, Impl, C-myc, RalA, p62, Survivin, and CyclinD1, were significantly different in various stages of esophageal cancer, which showed that autoantibodies may be involved in the pathogenesis and progression of esophageal cancer. All in all, this study further supports our previous hypothesis that a combination of antibodies might acquire higher sensitivity for the diagnosis of certain types of cancer. A customized mini-array of multiple carefully-selected TAAs is able to enhance autoantibody detection in the immunodiagnosis of esophageal cancer and autoantibodies to TAAs might be reference indicators of clinical stage.

Molecular characterization of the sans fille gene in Antheraea pernyi: A highly conserved gene during the evolution of animals

Ya-Jie Li,Rui Mi,Nan Meng,Zhi-Xin Wen,Xue-Jun Li,Mo Chen,Yan-Qun Liu,Shu-Ying Li 한국응용곤충학회 2013 Journal of Asia-Pacific Entomology Vol.16 No.3

Sans-fille (SNF) is the Drosophila homologue ofmammalian general splicing factors U1A and U2B″, and plays an important role in sex determination in Drosophilamelanogaster. In this study, the snf gene fromAntheraea pernyi (Lepidoptera: Saturniidae), an economically important insect, was isolated and characterized. The obtained 925 bp cDNA sequence contains an open reading frame of 669 bp encoding a polypeptide of 222 amino acids,showing 78% sequence identity to that from D. melanogaster. A database search revealed that SNF protein homologs are present in many animals, including invertebrates and vertebrates, with more than 70% amino acid sequence identities, suggesting that they were highly conserved during the evolution of animals. Phylogenetic analysis revealed that A. pernyi SNF was closely related to Bombyx mori SNF. Quantitative real-time PCR (qRT-PCR) analysis showed that the A. pernyi snf gene was transcribed during five larval developmental stages,and in six tested tissues (ovaries, testes, silk glands, fat body, integument, and hemolymph),with the most abundance determined in the gonads (ovaries or testes). Investigation of expression changes throughout embryonic development indicated that A. pernyi snfmRNAwas expressed at a lowlevel fromdays 0 to 4, and reached amaximum level at day 10, but decreased to a low level before hatching. These results suggest that the product of the snf gene may play important roles in the development of A. pernyi.

Epidemiological Patterns of Cancer Incidence in Southern China: Based on 6 Population-based Cancer Registries

Liu, Jie,Yang, Xu-Li,Li, Ai,Chen, Wan-Qing,Ji, Lu,Zhao, Jun,Yan, Wei,Chen, Yi-Ying,Zhu, Li-Ping Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.3

Background: The epidemiological patterns of cancer incidence have been investigated widely in western countries. Nevertheless, information is quite limited in Jiangxi province, southern China. Materials and Methods: All data were reported by 6 population-based cancer registries in Jiangxi Province. The results were presented as incidence rates of cases by site (ICD-10), sex, crude rate (CR), age-standardized rates (ASRs) and truncated incidence rate (TR) per 100,000 person-years, using the direct method of standardization to the world population. Results: 8,765 new cancer cases were registered in our study during the period 2009-2011. Diagnosis of cancer was based on histopathology in 61.0%, clinical or radiology findings in 4.87% and death certificate only (DCO) in 3.0% of the cases. The median age at diagnosis was 62.0 years (mean, 61; standard deviation, 15). The ASRs were 170.8 per 100,000 for men and 111.2 for women. The ASRs for all invasive cancers from the urban areas (145.7 per 100,000) was higher than that of rural areas (137.1). Incidence rates for lung cancer were higher in rural (35.8) than in urban areas (27.0). Similarly, relatively high rates were observed for stomach cancer in rural (20.1) relative to urban areas (15.5). Conclusions: Our results reveal that the most common cancers were breast and lung in women and lung and liver in men. Interestingly, this study suggested a higher incidence rates for lung and stomach cancer in rural males than in urban population, which may suggest other potential causes, such as over-consumption of smoked meats and high prevalence of Helicobacter pylori infection, respectively. Public education and the promotion of healthy lifestyles should be actively carried out.

Combined Assessment of Serum Alpha-Synuclein and Rab35 is a Better Biomarker for Parkinson’s Disease

Hung-Li Wang,Chin-Song Lu,Tu-Hsueh Yeh,Yu-Ming Shen,Yi-Hsin Weng,Ying-Zu Huang,Rou-Shayn Chen,Yu-Chuan Liu,Yi-Chuan Cheng,Hsiu-Chen Chang,Ying-Ling Chen,Yu-Jie Chen,Yan-Wei Lin,Chia Chen Hsu,Huang-Li 대한신경과학회 2019 Journal of Clinical Neurology Vol.15 No.4

Background and Purpose It is essential to develop a reliable predictive serum biomarker for Parkinson’s disease (PD). Te accumulation of alpha-synuclein (αSyn) and up-regulated expression of Rab35 participate in the etiology of PD. Te purpose of this investigation was to determine whether the combined assessment of serum αSyn and Rab35 is a useful predictive biomarker for PD. Methods Serum levels of αSyn or Rab35 were determined in serum samples from 59 sporadic PD patients, 19 progressive supranuclear palsy (PSP) patients, 20 multiple system atrophy (MSA) patients, and 60 normal controls (NC). Receiver operating characteristics (ROC) curves were calculated to determine the diagnostic accuracy of αSyn or/and Rab35 in discriminating PD patients from NC or atypical parkinsonian patients. Results The levels of αSyn and Rab35 were increased in PD patients. The serum level of Rab35 was positively correlated with that of αSyn in PD patients. Compared to analyzing αSyn or Rab35 alone, the combined analysis of αSyn and Rab35 produced a larger area under the ROC curve and performed better in discriminating PD patients from NC, MSA patients, or PSP patients. When age was dichotomized at 55, 60, 65, or 70 years, the combined assessment of αSyn and Rab35 for classifying PD was better in the group below the cutof age than in the group above the cutof age. Conclusions Combined assessment of serum αSyn and Rab35 is a better biomarker for discriminating PD patients from NC or atypical parkinsonian patients, and is a useful predictive biomarker for younger sporadic PD patients.

High-Definition Computed Tomography for Coronary Artery Stent Imaging: a Phantom Study

Wen Jie Yang,Ke Min Chen,Li Fang Pang,Ying Guo,Jian Ying Li,Huang Zhang,Zi Lai Pan 대한영상의학회 2012 Korean Journal of Radiology Vol.13 No.1

Objective: To assess the performance of a high-definition CT (HDCT) for imaging small caliber coronary stents (≤ 3 mm) by comparing different scan modes of a conventional 64-row standard-definition CT (SDCT). Materials and Methods: A cardiac phantom with twelve stents (2.5 mm and 3.0 mm in diameter) was scanned by HDCT and SDCT. The scan modes were retrospective electrocardiography (ECG)-gated helical and prospective ECG-triggered axial with tube voltages of 120 kVp and 100 kVp, respectively. The inner stent diameters (ISD) and the in-stent attenuation value (AVin-stent) and the in-vessel extra-stent attenuation value (AVin-vessel) were measured by two observers. The artificial lumen narrowing (ALN = [ISD - ISDmeasured]/ISD) and artificial attenuation increase between in-stent and in-vessel (AAI = AVin-stent - AVin-vessel) were calculated. All data was analyzed by intraclass correlation and ANOVA-test. Results: The correlation coefficient of ISD, AVin-vessel and AVin-stent between the two observers was good. The ALNs of HDCT were statistically lower than that of SDCT (30 ± 5.7% versus 35 ± 5.4%, p < 0.05). HDCT had statistically lower AAI values than SDCT (15.7 ± 81.4 HU versus 71.4 ± 90.5 HU, p < 0.05). The prospective axial dataset demonstrated smaller ALN than the retrospective helical dataset on both HDCT and SDCT (p < 0.05). Additionally, there were no differences in ALN between the 120 kVp and 100 kVp tube voltages on HDCT (p = 0.05). Conclusion: High-definition CT helps improve measurement accuracy for imaging coronary stents compared to SDCT. HDCT with 100 kVp and the prospective ECG-triggered axial technique, with a lower radiation dose than 120 kVp application, may be advantageous in evaluating coronary stents with smaller calibers (≤ 3 mm). Objective: To assess the performance of a high-definition CT (HDCT) for imaging small caliber coronary stents (≤ 3 mm) by comparing different scan modes of a conventional 64-row standard-definition CT (SDCT). Materials and Methods: A cardiac phantom with twelve stents (2.5 mm and 3.0 mm in diameter) was scanned by HDCT and SDCT. The scan modes were retrospective electrocardiography (ECG)-gated helical and prospective ECG-triggered axial with tube voltages of 120 kVp and 100 kVp, respectively. The inner stent diameters (ISD) and the in-stent attenuation value (AVin-stent) and the in-vessel extra-stent attenuation value (AVin-vessel) were measured by two observers. The artificial lumen narrowing (ALN = [ISD - ISDmeasured]/ISD) and artificial attenuation increase between in-stent and in-vessel (AAI = AVin-stent - AVin-vessel) were calculated. All data was analyzed by intraclass correlation and ANOVA-test. Results: The correlation coefficient of ISD, AVin-vessel and AVin-stent between the two observers was good. The ALNs of HDCT were statistically lower than that of SDCT (30 ± 5.7% versus 35 ± 5.4%, p < 0.05). HDCT had statistically lower AAI values than SDCT (15.7 ± 81.4 HU versus 71.4 ± 90.5 HU, p < 0.05). The prospective axial dataset demonstrated smaller ALN than the retrospective helical dataset on both HDCT and SDCT (p < 0.05). Additionally, there were no differences in ALN between the 120 kVp and 100 kVp tube voltages on HDCT (p = 0.05). Conclusion: High-definition CT helps improve measurement accuracy for imaging coronary stents compared to SDCT. HDCT with 100 kVp and the prospective ECG-triggered axial technique, with a lower radiation dose than 120 kVp application, may be advantageous in evaluating coronary stents with smaller calibers (≤ 3 mm).

Ontogenetic Expression of Lpin2 and Lpin3 Genes and Their Associations with Traits in Two Breeds of Chinese Fat-tailed Sheep

Jiao, Xiao-Li,Jing, Jiong-Jie,Qiao, Li-Ying,Liu, Jian-Hua,Li, Liu-An,Zhang, Jing,Jia, Xia-Li,Liu, Wen-Zhong Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.3

Lipins play dual function in lipid metabolism by serving as phosphatidate phosphatase and transcriptional co-regulators of gene expression. Mammalian lipin proteins consist of lipin1, lipin2, and lipin3 and are encoded by their respective genes Lpin1, Lpin2, and Lpin3. To date, most studies are concerned with Lpin1, only a few have addressed Lpin2 and Lpin3. Ontogenetic expression of Lpin2 and Lpin3 and their associations with traits would help to explore their molecular and physiological functions in sheep. In this study, 48 animals with an equal number of males and females each for both breeds of fat-tailed sheep such as Guangling Large Tailed (GLT) and Small Tailed Han (STH) were chosen to evaluate the ontogenetic expression of Lpin2 and Lpin3 from eight different tissues and months of age by quantitative real-time polymerase chain reaction (PCR). Associations between gene expression and slaughter and tail traits were also analyzed. The results showed that Lpin2 mRNA was highly expressed in perirenal and tail fats, and was also substantially expressed in liver, kidney, reproductive organs (testis and ovary), with the lowest levels in small intestine and femoral biceps. Lpin3 mRNA was prominently expressed in liver and small intestine, and was also expressed at high levels in kidney, perirenal and tail fats as well as reproductive organs (testis and ovary), with the lowest level in femoral biceps. Global expression of Lpin2 and Lpin3 in GLT both were significantly higher than those in STH. Spatiotemporal expression showed that the highest levels of Lpin2 expression occurred at 10 months of age in two breeds of sheep, with the lowest expression at 2 months of age in STH and at 8 months of age in GLT. The greatest levels of Lpin3 expression occurred at 4 months of age in STH and at 10 months of age in GLT, with the lowest expression at 12 months of age in STH and at 8 months of age in GLT. Breed and age significantly influenced the tissue expression patterns of Lpin2 and Lpin3, respectively, and sex significantly influenced the spatiotemporal expression patterns of Lpin3. Meanwhile, Lpin2 and Lpin3 mRNA expression both showed significant correlations with slaughter and tail traits, and the associations appear to be related with the ontogenetic expression as well as the potential functions of lipin2 and lipin3 in sheep.