A Highly Active Alpha Amylase from Bacillus Licheniformis: Directed Evolution, Enzyme Characterization and Structural Analysis

( Yi Han Liu ),( Shu Ai Fan ),( Xiao Guang Liu ),( Zhi Meng Zhang ),( Jian Ling Wang ),( Zheng Xiang Wang ),( Fu Ping Lu ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.7

The stability of Bacillus licheniformis alpha-amylase (BLA) under acid condition was enhanced through direct evolution using the error-prone polymerase chain reaction. One beneficial mutation site, H281I, was obtained in BLA. The specific activity of H281I was 161/352 U/mg, which was 62.6/27.5% higher than that of the wild-type (WT) (99/276 U/mg) at pH 4.5/6.5 and 95°C. The pH optimum for H281I was decreased about 1 unit, whereas no significant changes of optimum temperature and thermostability were observed compared with the wild type (WT). The kcat/Km value of H281I was 1.7-/1.4-fold higher at pH 4.5/6.5, respectively, than that of WT. The structure model analysis indicated that the H281I mutation altered the predicted interaction between the amino acid residues at 281 and 273, thus creating a conducive local environment for substrate binding, as reflected by its decreased Km, and consequently increased the specific activity.

Pulmonary Lobectomy Combined with Pulmonary Arterioplasty by Complete Video-assisted Thoracic Surgery in Patients with Lung Cancer

Yu, Da-Ping,Han, Yi,Zhao, Qiu-Yue,Liu, Zhi-Dong Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.10

Objective: To explore the feasibility of pulmonary lobectomy combined with pulmonary arterioplasty by complete video-assisted thoracic surgery (VATS) in patients with lung cancer, and summarize its surgical methods. Materials and Methods: Twenty-one patients with lung cancer in Beijing Chest Hospital Affiliated to Capital Medical University from Feb., 2010 to Jun., 2013 were selected, males and females accounting for 15 and 6 cases, respectively. Ten underwent right upper lobectomy, 5 right lower lobectomy, 4 left upper lobectomy (in which left upper sleeve lobectomy was conducted for 2) and 2 left lower lobectomy. At the same time, local resection of pulmonary arterioplasty was performed for 12 patients, and sleeve resection of pulmonary arterioplasty for 9. Results: Twenty-one patients recovered well after surgery. Thoracic drainage tube was maintained for 3-8 days, with an average of 4.9 days, and hospital stays were 8-15 days, with an average of 11 days. There were no deaths in the perioperative period, and the complications like pulmonary embolism, bronchopleural fistula, chest infection and pulmonary atelectasis did not occur after surgery. Conclusions: Performance of pulmonary lobectomy and pulmonary arterioplasty together by complete VATS is a safe and effective surgical method, which can expand the indications of patients with lung cancer undergoing thoracoscopic pulmonary lobectomy, and make more patients profit from such minimally invasive treatment.

CD3⁺ CD4⁺ and CD3⁺ CD8⁺ Lymphocyte Subgroups and their Surface Receptors NKG2D and NKG2A in Patients with Non-small Cell Lung Cancer

Yu, Da-Ping,Han, Yi,Zhao, Qiu-Yue,Liu, Zhi-Dong Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

Background: To explore the prevalence of lymphocyte subgroups $CD3^+$ $CD4^+$ and $CD3^+$ $CD8^+$ and their surface receptors NKG2D and NKG2A in patients with non-small cell lung cancer (NSCLC). Materials and Methods: A total of 40 patients with NSCLC were divided into different groups according to different clinical factors (TNM staging, pathological patterns and genders) for assessment of relations with $CD3^+$ $CD4^+$ and $CD3^+$ $CD8^+$ and the surface receptors NKG2D and NKG2A of T lymphocytes in peripheral blood by flow cytometry. Results: Patients in the advanced group had evidently lower levels of $CD3^+$ $CD4^+$ but markedly higher levels of $CD3^+$ $CD8^+$ in peripheral blood than those with early lesions (p<0.05). In addition, NSCLC patients in the advanced group had obviously higher $CD3^+$ $CD4^+$ NKG2D and $CD3^+$ $CD8^+$ NKG2A expression rates but lower $CD3^+$ $CD4^+$ NKG2A and $CD3^+$ $CD8^+$ NKG2D expression rates (p<0.05). However, there were no significant differences between NSCLC patients with different genders and pathological patterns in expression levels of lymphocyte subgroups $CD3^+$ $CD4^+$ and $CD3^+$ $CD8^+$ and their surface receptors NKG2D and NKG2A. Conclusions: Unbalanced expression of surface receptors NKG2D and NKG2A in $CD3^+$ $CD4^+$ and $CD3^+$ $CD8^+$ lymphocytes may be associated with a poor prognosis, greater malignancy and immunological evasion by advanced cancers, related to progression of lung cancer.

The first mitochondrial genome for Brahmin moths (Brahmaeidae) and implications for the higher phylogeny of Bombycoidea

Hong-Li Zhang,Run-Mei Wang,Zhi-Ping Han,Yong-Fang Zhang,Jian-Xia Liu 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.2

Bombycoidea comprises 10 families and 4723 species, and the phylogenetic relationships among families are still in debate. In this study, we have determined the complete mitochondrial genome (mitogenome) of Brahmaea porphyria. The 15,429-bp mitogenome contains a common set of 37 mitochondrial genes including 13 proteincoding genes, 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs) and an inferred control region, and shares the conserved gene rearrangement (trnM-trnI-trnQ) in most ditrysian mitogenomes. Moreover, we analysed the secondary structure for all the tRNA genes of B. porphyria and the preference of codon usage in the PCGs of B. porphyria. The putative 373-bp control region (CR) possesses three types of conserved elements, including ATAGA, Ploy-T stretch, and microsatellite-like elements. A phylogenetic analysis among available Bombycoidea mitogenomes using the concatenated 37 mitochondrial genes appears to support the hypothesis of (Sphingidae+Bombycidae)+Saturniidae and the relatively basal phylogenetic position of Brahmaeidae within Bombycoidea.

Nomogram for predicting overall survival in children with neuroblastoma based on SEER database

Song-Wu Liang,Gang Chen,Yi-Ge Luo,Peng Chen,Jin-Han Gu,Qiong-Qian Xu,Yi-Wu Dang,Li-Ting Qin,Hui-Ping Lu,Wen-Ting Huang,Zhi-Guang Huang,Li Gao,Jia-Bo Chen 대한외과학회 2020 Annals of Surgical Treatment and Research(ASRT) Vol.99 No.2

Purpose: This study was performed to establish and validate a nomogram for predicting the overall survival in children with neuroblastoma. Methods: The latest clinical data of neuroblastoma in Surveillance, Epidemiology, and End Results (SEER) database was extracted from 2000 to 2016. The cases included were randomly divided into training and validation cohorts. The survival curves were drawn with a Kaplan-Meier estimator to investigate the influences of certain single factors on overall survival. Also, least absolute shrinkage and selection operator regression was applied to further select the prognostic variables for neuroblastoma. Additionally, receiver operating characteristic (ROC) curves and calibration curves were used to evaluate the accuracy of the nomogram. Results: In total, 1,262 patients were collected and 8 independent prognostic factors were achieved, including patients’ age, sex, race, tumor grade, radiotherapy, chemotherapy, tumor site, and tumor size. Then we constructed a nomogram by using the data of the training cohort with 886 cases. Subsequently, the nomogram was validated internally and externally with 886 and 376 cases, respectively. The internal validation revealed that the area under the curves (AUC) of ROC curves of 1-, 3-, and 5-year overall survival were 0.69, 0.78, and 0.81, respectively. Accordingly, the external validation also showed that the AUC of 1-, 3-, and 5-year overall survival were all ≥0.69. Both methods of validation demonstrated that the predictive calibration curves were consistent with standard curves. Conclusion: The nomogram possess the potential to be a new tool in predicting the survival rate of neuroblastoma patients.

Characterization of proteolysis in muscle tissues of sea cucumber Stichopus japonicus

Chen-Chen Zhao,Yang Yang,Hai-tao Wu,Zhi-Mo Zhu,Yue Tang,Cui-Ping Yu,Na Sun,Qiang Lv,Jia-Run Han,Ao-Ting Li,Jia-Nan Yan,Yue Cha 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.6

The proteolysis in muscle tissues of sea cucumber Stichopus japonicus (sjMTs) was characterized. The proteins from sjMTs were primarily myosin heavy chains (MHCs), paramyosin (Pm), and actin (Ac) having a molecular mass of approximately 200, 98, and 42 kDa, respectively. Based on SDS-PAGE analysis and quantification of trichloroacetic acid (TCA)-soluble peptides released, degradation of muscle proteins from sjMTs was favorable at pH 5 and 50°C. Proteolysis of MHCs was mostly inhibited by cysteine protease inhibitors, including trans-epoxysuccinyl-L-leucyl-amido (4- guanidino) butane (E-64) and antipain (AP). E-64 and AP completely inhibited the degradation of Pm and Ac, while iodoacetic acid showed a partially inhibitory effect. These results indicated that the proteolysis of sjMTs was mainly attributed to cysteine proteases. Avoidance of setting the tissues at 40–50oC and slightly acidic condition and inhibition of cysteine proteases are helpful for decreasing sea cucumber autolysis.

ZNF488 Enhances the Invasion and Tumorigenesis in Nasopharyngeal Carcinoma Via the Wnt Signaling Pathway Involving Epithelial Mesenchymal Transition

Dan Zong,Li Yin,Qian Zhong,Wen-jie Guo,Jian-hua Xu,Ning Jiang,Zhi-rui Lin,Man-zhi Li,Ping Han,Lin Xu,Xia He,Mu-sheng Zeng 대한암학회 2016 Cancer Research and Treatment Vol.48 No.1

Purpose The purpose of this study was to investigate the function of Zinc finger protein 488 (ZNF488) in nasopharyngeal carcinoma (NPC). Materials and Methods The endogenous expression of ZNF488 in NPC tissues, normal nasopharyngeal epithelium tissues and NPC cell lines were detected by quantitative reverse transcription polymerase chain reaction. ZNF488 over-expressing and knock-down NPC cell line models were estab- lished through retroviral vector pMSCV mediated over-expression and small interfering RNA (siRNA) mediated knock-down. The invasion and migration capacities were evaluated by wound healing and transwell invasion assays in ZNF488 over-expressing and control cell lines. Soft-agar colony formation and a xenograft experiment were performed to study tumorigenic ability in vitro and in vivo. Immunofluorescence and western blotting analysis were used to examine protein changes followed by ZNF488 over-expression. Microarray analysis was performed to explore gene expression profilings, while luciferase reporter assay to evaluate the transcriptive activity of Tcf/Lef. Results ZNF488 was over-expressed in NPC tissues compared with normal tissues, especially higher in 5-8F and S18, which are well-established high metastatic NPC clones. Functional studies indicate that over-expression of ZNF488 provokes invasion, whereas knock-down of ZNF488 alleviates invasive capability. Moreover, over-expression of ZNF488 promotes NPC tumor growth both in vitro and in vivo. Our data further show that over-expression of ZNF488 induces epithelial mesenchymal transition (EMT) by activating the WNT/β -catenin signaling pathway. Conclusion Our data strongly suggest that ZNF488 acts as an oncogene, promoting invasion and tumorigenesis by activating the Wnt/β -catenin pathway to induce EMT in NPC.

Circular RNA cFAM210A, degradable by HBx, inhibits HCC tumorigenesis by suppressing YBX1 transactivation

Yu Jian,Li Wen,Hou Guo-jun,Sun Da-peng,Yang Yuan,Yuan Sheng-xian,Dai Zhi-hui,Yin Hao-zan,Sun Shu-han,Huang Gang,Zhou Wei-ping,Yang Fu 생화학분자생물학회 2023 Experimental and molecular medicine Vol.55 No.-

Hepatitis B protein x (HBx) has been reported to promote tumorigenesis in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), but the mechanism awaits further investigation. In this study, we found that cFAM210A (a circular RNA derived from the third exon of transcript NM_001098801 of the FAM210A gene; CircBase ID: hsa_circ_0003979) can be silenced by HBx. cFAM210A expression was downregulated and negatively correlated with tumorigenesis in patients with HBV-related HCC. Furthermore, cFAM210A reduced the proliferation, stemness, and tumorigenicity of HCC cells. Mechanistically, HBx increased the N6-methyladenosine (m6A) level of cFAM210A by promoting the expression of RBM15 (an m6A methyltransferase), thus inducing the degradation of cFAM210A via the YTHDF2-HRSP12-RNase P/MRP pathway. cFAM210A bound to YBX1 and inhibited its phosphorylation, suppressing its transactivation function toward MET. These findings suggest the important role of circular RNAs in HBx-induced hepatocarcinogenesis and identify cFAM210A a potential target in the prevention and treatment of HBV-related HCC.