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.

Expression and Prognostic Implications of FOXO3a and Ki67 in Lung Adenocarcinomas

Liu, Hong-Bin,Gao, Xiang-Xiang,Zhang, Qing,Liu, Jian,Cui, Yuan,Zhu, Yan,Liu, Yi-Fei Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.4

To investigate the significance of FOXO3a and Ki67 in human lung adenocarcinomas. Envision immunohistochemical staining and Western blotting were used to examine the protein expression of FOXO3a in 127 cases of human lung adenocarcinoma specimens. The positive rate in lung adenocarcinoma (55.9%) was lower than that in normal tissues (80%). We found that the expression of FOXO3a was closely related with the degree of differentiation, TNM staging, lymph node metastasis and survival. In addition, significant differences in the different pathological types of lung adenocarcinoma cases (P<0.01). The FOXO3a positive rate of the acini as the main type (APA) (86.7%) and the lepidic as the main type (LPA) (82.4%) was higher than the solid as the main type (SPA) (50.0%), the papilla as the main type (PPA) (42.9%) and the micropapilla as the main type (MPA) (9.4%). Moreover, the expression of FOXO3a was negatively related with Ki67 expression. Our results suggested that the expression of FOXO3a is closely correlated with the aggressiveness of lung adenocarcinoma. It was indicated that disregulation of FOXO3a might play key roles in the occurrence and development of lung a denocarcinoma and joint detection of the two markers might play an important role in diagnosing tumors.

Treatment of bladder cancer by geoinspired synthetic chrysotile nanocarrier-delivered circPRMT5 siRNA

Chunping Yu,Yi Zhang,Ning Wang,Wensu Wei,Ke Cao,Qun Zhang,Peiying Ma,Dan Xie,Pei Wu,Biao Liu,Jiahao Liu,Wei Xiang,Xing Hu,Xuewen Liu,Jianfei Xie,Jin Tang,Zhi Long,Long Wang,Hongliang Zeng,Jianye Liu 한국생체재료학회 2022 생체재료학회지 Vol.26 No.1

Background: Circular RNAs (circRNAs) have important functions in many fields of cancer biology. In particular, we previously reported that the oncogenic circRNA, circPRMT5, has a major role in bladder cancer progression. Therapy based on circRNAs have good prospects as anticancer strategies. While anti-circRNAs are emerging as therapeutics, the specific in vivo delivery of anti-circRNAs into cancer cells has not been reported and remains challenging. Methods: Synthesized chrysotile nanotubes (SCNTs) with a relatively uniform length (~ 200 nm) have been designed to deliver an siRNA against the oncogenic circPRMT5 (si-circPRMT5) inhibit circPRMT5. In addition, the antitumor effects and safety evaluation of SCNTs/si-circPRMT5 was assessed with a series of in vitro and in vivo assays. Results: The results showed that SCNTs/si-circPRMT5 nanomaterials prolong si-circPRMT5’s half-life in circulation, enhance its specific uptake by tumor cells, and maximize the silencing efficiency of circPRMT5. In vitro, SCNTs encapsulating si-circPRMT5 could inhibit bladder cancer cell growth and progression. In vivo, SCNTs/si-circPRMT5 inhibited growth and metastasis in three bladder tumor models (a subcutaneous model, a tail vein injection lung metastatic model, and an in situ model) without obvious toxicities. Mechanistic study showed that SCNTs/sicircPRMT5 regulated the miR-30c/SNAIL1/E-adherin axis, inhibiting bladder cancer growth and progression. Conclusion: The results highlight the potential therapeutic utility of SCNTs/si-circPRMT5 to deliver si-circPRMT5 to treat bladder cancer.

Distributed Space-Time Coding for Full-Duplex Asynchronous Cooperative Communications

Yi Liu,Xiang-Gen Xia,Hailin Zhang IEEE 2012 IEEE TRANSACTIONS ON WIRELESS COMMUNICATIONS Vol.11 No.7

<P>In this paper, we propose two distributed linear convolutional space-time coding (DLC-STC) schemes for full-duplex (FD) asynchronous cooperative communications. The DLC-STC Scheme 1 is for the case of the complete loop channel cancellation, which achieves the full asynchronous cooperative diversity. The DLC-STC Scheme 2 is for the case of the partial loop channel cancellation and amplifying, where some loop signals are used as the self-coding instead of treated as interference to be directly cancelled. We show this scheme can achieve full asynchronous cooperative diversity. We then evaluate the performance of the two schemes when loop channel information is not accurate and present an amplifying factor control method for the DLC-STC Scheme 2 to improve its performance with inaccurate loop channel information. Simulation results show that the DLC-STC Scheme 1 outperforms the DLC-STC Scheme 2 and the delay diversity scheme if perfect or high quality loop channel information is available at the relay, while the DLC-STC Scheme 2 achieves better performance if the loop channel information is imperfect.</P>

Fresh Washed Microbiota Transplantation Alters Gut Microbiota Metabolites to Ameliorate Sleeping Disorder Symptom of Autistic Children

Liu Nai-Hua,Liu Hong-Qian,Zheng Jia-Yi,Zhu Meng-Lu,Wu Li-Hao,Pan Hua-Feng,He Xing-Xiang 한국미생물학회 2023 The journal of microbiology Vol.61 No.8

Accumulating studies have raised concerns about gut dysbiosis associating autism spectrum disorder (ASD) and its related symptoms. However, the effect of gut microbiota modification on the Chinese ASD population and its underlying mechanism were still elusive. Herein, we enrolled 24 ASD children to perform the first course of fresh washed microbiota transplantation (WMT), 18 patients decided to participate the second course, 13 of which stayed to participate the third course, and there were 8 patients at the fourth course. Then we evaluated the effects of fresh WMT on these patients and their related symptoms. Our results found that the sleeping disorder symptom was positively interrelated to ASD, fresh WMT significantly alleviated ASD and its sleeping disorder and constipation symptoms. In addition, WMT stably and continuously downregulated Bacteroides/ Flavonifractor/Parasutterella while upregulated Prevotella_9 to decrease toxic metabolic production and improve detoxification by regulating glycolysis/myo-inositol/D-glucuronide/D-glucarate degradation, L-1,2-propanediol degradation, fatty acid β-oxidation. Thus, our results suggested that fresh WMT moderated gut microbiome to improve the behavioral and sleeping disorder symptoms of ASD via decrease toxic metabolic production and improve detoxification. Which thus provides a promising gut ecological strategy for ASD children and its related symptoms treatments.

Chemotherapy and Late Course Three Dimensional Conformal Radiotherapy for Treatment of Patients with Stage III Non-small Cell Lung Cancer

Liu, Yang-Chen,Zhou, Shao-Bing,Gao, Fei,Ye, Hong-Xun,Zhao, Ying,Yi, Xiao-Xiang,Huang, Xin-En,Xiang, Jin Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.4

Objective: To compare the efficacy and complications of chemotherapy and late course three-dimensional conformal radiotherapy (3DCRT) in treating patients with stage III non-small cell lung cancer (NSCLC). Patients and Methods: All patients were divided into two groups: to receive chemotherapy and late course 3DCRT (3DCRT group), or chemotherapy and conventional fraction radiation (control group). In the 3DCRT-group, patients were given 6~15 MV X-rays with a total dose of 40 Gy, followed by 3DCRT, 2.5 Gy~3.0 Gy per fraction, 1 fraction/every day, total 68 Gy~70 Gy; in the control group, with conventional fraction radiation the total dose was 64~66 Gy. The chemotherapy regimen in both cases was EP (VP-16 and DDP). Results: Sixty four patients with stage III NSCLC were divided into two groups: 32 patients into 3DCRT, 32 into the control group. One and 2-year survival rates in 3DCRT and control group were 87.5%, 56.3%mad 65.6%, 34.4%, respectively (P<0.05); local control rates were 90.6%, 81.3% and 65.6%, 53.1%, respectively (P<0.05). Conclusion: Chemotherapy and late course 3DCRT is associated with improved survival rate in patients with stage III NSCLC with good tolerability.

Effects of Alpha 1- and Alpha 2-Adrenoreceptor Stimulation on Galanin mRNA Expression in Primary Cultured Superior Cervical Ganglion Neurons

( Yi Xing ),( Xiu Ying Chen ),( Zhen Liu ),( Hao Li ),( Hua Xiang Liu ),( Zhen Zhong Li ) 한국응용약물학회 2011 Biomolecules & Therapeutics(구 응용약물학회지) Vol.19 No.3

Galanin (Gal) is a 29-amino-acid neuropeptide which is expressed in superior cervical ganglion (SCG) neurons and plays a trophic role in the adult animal and acts as an inhibitory modulator of cholinergic and noradrenergic neurotransmission. Whether activation or inhibition of alpha-adrenoreceptors infl uences Gal mRNA expression in SCG neurons remains unknown. Here, we have evaluated the possible regulation of Gal mRNA expression with acute (4 h) and chronic (4 days) stimulation of alpha 1- and alpha 2-adrenoreceptor agonists or antagonists in primary cultured SCG neurons. The results showed that the amount of Gal mRNA expression in cultured SCG neurons increased significantly after chronic stimulation with alpha 2-adrenoreceptor antagonist yohimbine compared with control SCG neurons at the same time point, whereas the amount of Gal mRNA expression decreased significantly after chronic stimulation with alpha 2-adrenoreceptor agonist clonidine as compared with that in control group. All these effects were not dose-dependent on the administration of alpha 2-adrenoreceptor agonist clonidine or alpha 2-adrenoreceptor antagonist yohimbine. Alpha 1-adrenoreceptor agonist phenylephrine or antagonist prazosin chronic stimulation did not have effects on Gal mRNA expression. Acute exposure of these agents did not have effects on Gal mRNA expression. The present study showed that Gal may be regulated by activation or inhibition of alpha 2-adrenoreceptors, but not alpha 1-adrenoreceptors in sympathetic neurons.

Cytotoxic T Lymphocytes Elicited by Dendritic Cell-Targeted Delivery of Human Papillomavirus Type-16 E6/E7 Fusion Gene Exert Lethal Effects on CaSki Cells

Wu, Xiang-Mei,Liu, Xing,Jiao, Qing-Fang,Fu, Shao-Yue,Bu, You-Quan,Song, Fang-Zhou,Yi, Fa-Ping Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety and non human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeutic vaccine, although none have so far been reported as effective. Since tumor cells consistently express the two proteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DC vaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene of HPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro. Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 to prepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11c was assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined by DAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro, and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to induce specific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells were significantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccine modified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as a new strategy for biological treatment of cervical cancer.

Nb reinforced Fe-Mn-Si shape memory alloy composite coating fabricated by laser cladding on 304 stainless steel surface

Xiaochen Liu,Peng Xu,Like Yao,Xiang Li,Chi Pang,Long Yang,Yi Liang 대한기계학회 2022 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.36 No.10

Laser cladding technology was adapted to fabricate Fe-based shape memory alloy/Nb composite coatings with 5 wt. % Nb doping amounts on 304 stainless steel to promote the microhardness and wear resistance. The SEM, XRD, optical profilometer and electrochemical workstation were used to characterize micromorphology, phase and microstructure. Meanwhile, wear and anti-resistance ability were detected. The results showed that Nb, NbC, ε-martensite, α’-martensite and γ-austenite were found in SMA/Nb coating. The microhardness was enhanced because of the solid solution strengthening and second phase strengthening. Also, the shape recovery rate and anti-corrosion property of the SMA/Nb coating are improved as well.

Clinical Safety and Efficacy of Kanglaite^® (Coix Seed Oil) Injection Combined with Chemotherapy in Treating Patients with Gastric Cancer

Zhan, Yi-Ping,Huang, Xin-En,Cao, Jie,Lu, Yan-Yan,Wu, Xue-Yan,Liu, Jin,Xu, Xia,Xiang, Jin,Ye, Li-Hong Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.10

Objective: To observe efficacy and side effects, as well as the impact on quality of life, of Kanglaite$^{(R)}$ (Coix Seed Oil) injections combined with chemotherapy in the treatment of advanced gastric cancer patients. Method: A consecutive cohort of 60 patients were divided into two groups: the experimental group receiving Kanglaite$^{(R)}$ Injection combined with chemotherapy and the control group with chemotherapy alone. After more than two courses of treatment, efficacy, quality of life and side effects were evaluated. Results: The response rate and KPS score of experimental group were significantly improved as compared with those of the control group (P<0.05). In addition, gastrointestinal reactions and bone marrow suppression were significantly lower than in the control group (P<0.05). Conclusions: Kanglaite$^{(R)}$ Injection enhanced efficacy and reduced the side effects of chemotherapy, improving quality of life of gastric cancer patients; use of Kanglaite$^{(R)}$ injections deserves to be further investigated in randomized control clinical trails.