Purification and Structural Analysis of Surfactin Produced by Endophytic Bacillus subtilis EBS05 and its Antagonistic Activity Against Rhizoctonia cerealis

Cai-Yi Wen,Shun-Shan Shen,Zhi-Gang Yin,Kai-Xuan Wang,Jian-Guang Chen 한국식물병리학회 2011 Plant Pathology Journal Vol.27 No.4

Bacillus subtilis EBS05, an endophytic bacteria strain isolated from a medicinal plant Cinnamomum camphor,can produce antagonistic compounds that effectively inhibit plant pathogenic fungi. The greenhouse experiments showed that wheat sharp eyespot disease (WSED)was reduced by 91.2%, 88.2% and 43.0% after the treatment with fermentation broth, bacteria-free filter and a fungicide fludioxonil, respectively. The culture broth of strain EBS05 can more effectively control WSED than can fludioxonil. The fermentation broth and bacteria-free filter ability to suppress WSED was not significantly different, suggesting that an active secreted substance played a major role in controlling WSED. Separation and purification of the active compounds was carried out by serial processes, including hydrochloric acid (pH 2.0) treatment, methanol extraction and Sephadex LH-20 column chromatography,silica gel column chromatography and reverse-phase high-pressure liquid chromatography (HPLC), respectively. The purified compounds, one of active peaks in the HPLC spectrum, were obtained from the collection. Analysis of the chemical structures by time-of-flight mass spectrometry (TOF-MS) and electrospray ionization mass spectrometry/mass spectrometry (ESI-MS/MS) showed that the active substances produced by the endophytic bacteria EBS05 are mixture of the β-hydroxy-C12~C15-Leu^7 surfactin A isomers with 1035.65Da, 1021.64 Da, 1007.63 Da and 993.65 Da molecular weights, respectively.

Purification and Structural Analysis of Surfactin Produced by Endophytic Bacillus subtilis EBS05 and its Antagonistic Activity Against Rhizoctonia cerealis

Wen, Cai-Yi,Yin, Zhi-Gang,Wang, Kai-Xuan,Chen, Jian-Guang,Shen, Shun-Shan The Korean Society of Plant Pathology 2011 Plant Pathology Journal Vol.27 No.4

Bacillus subtilis EBS05, an endophytic bacteria strain isolated from a medicinal plant Cinnamomum camphor, can produce antagonistic compounds that effectively inhibit plant pathogenic fungi. The greenhouse experiments showed that wheat sharp eyespot disease (WSED) was reduced by 91.2%, 88.2% and 43.0% after the treatment with fermentation broth, bacteria-free filter and a fungicide fludioxonil, respectively. The culture broth of strain EBS05 can more effectively control WSED than can fludioxonil. The fermentation broth and bacteria-free filter ability to suppress WSED was not significantly different, suggesting that an active secreted substance played a major role in controlling WSED. Separation and purification of the active compounds was carried out by serial processes, including hydrochloric acid (pH 2.0) treatment, methanol extraction and Sephadex LH-20 column chromatography, silica gel column chromatography and reverse-phase high-pressure liquid chromatography (HPLC), respectively. The purified compounds, one of active peaks in the HPLC spectrum, were obtained from the collection. Analysis of the chemical structures by time-of-flight mass spectrometry (TOF-MS) and electrospray ionization mass spectrometry/mass spectrometry (ESI-MS/MS) showed that the active substances produced by the endophytic bacteria EBS05 are mixture of the ${\beta}$-hydroxy-C12~C15-$Leu^7$ surfactin A isomers with 1035.65 Da, 1021.64 Da, 1007.63 Da and 993.65 Da molecular weights, respectively.

Phenotypic and Molecular Characteristics of Children with Progressive Familial Intrahepatic Cholestasis in South China

Wen Zhang,Ruizhu Lin,Zhikun Lu,Huiying Sheng,Yi Xu,Xiuzhen Li,Jing Cheng,Yanna Cai,Xiaojian Mao,Li Liu 대한소아소화기영양학회 2020 Pediatric gastroenterology, hepatology & nutrition Vol.23 No.6

Purpose: Progressive familial intrahepatic cholestasis (PFIC) is a rare genetic autosomal recessive disease caused by mutations in ATP8B1, ABCB11 or ABCB4. Mutational analysis of these genes is a reliable approach to identify the disorder. Methods: We collected and analyzed relevant data related to clinical diagnosis, biological investigation, and molecular determination in nine children carrying these gene mutations, who were from unrelated families in South China. Results: Of the nine patients (five males, four females) with PFIC, one case of PFIC1, four cases of PFIC2, and four cases of PFIC3 were diagnosed. Except in patient no. 8, jaundice and severe pruritus were the major clinical signs in all forms. γ-glutamyl transpeptidase was low in patients with PFIC1/PFIC2, and remained mildly elevated in patients with PFIC3. We identified 15 different mutations, including nine novel mutations (p.R470HfsX8, p.Q794X and p.I1170T of ABCB11 gene mutations, p.G319R, p.A1047P, p.G1074R, p.T830NfsX11, p.A1047PfsX8 and p.N1048TfsX of ABCB4 gene mutations) and six known mutations (p.G446R and p.F529del of ATP8B1 gene mutations, p.A588V, p.G1004D and p.R1057X of ABCB11 gene mutations, p.P479L of ABCB4 gene mutations). The results showed that compared with other regions, these three types of PFIC genes had different mutational spectrum in China. Conclusion: The study expands the genotypic spectrum of PFIC. We identified nine novel mutations of PFIC and our findings could help in the diagnosis and treatment of this disease.

Four-Hour Fasting for Semifluids and 2-Hour Fasting for Water Improves the Patient Experience of Esophagogastroduodenoscopy: A Randomized Controlled Trial

Cai Meng-Xi,Gao Ye,Li Li,Feng Wen,Wang Yi-Lin,Li Zhao-Shen,Xin Lei,Wang Luo-Wei 거트앤리버 소화기연관학회협의회 2023 Gut and Liver Vol.17 No.3

Background/Aims: We aimed to investigate the comfort, safety, and endoscopic visibility during esophagogastroduodenoscopy (EGD) afforded by a modified 4-hour semifluid and 2-hour water (“4+2”) fasting protocol. Methods: In this parallel group, endoscopist-blinded, randomized controlled trial, outpatients undergoing unsedated diagnostic EGD from 10:30 AM to 12:00 PM were randomly assigned to either a “4+2” protocol group or a conventional fasting group. The participants’ comfort during the fasting period and procedure was measured using the visual analog scale, and mucosal visibility was measured by endoscopists using the total visibility score. Satisfaction was defined as a visual analog scale score of ≤3. The primary outcome was the participants’ comfort during fasting. Results: One hundred and six and 108 participants were randomized to the “4+2” protocol and control groups, respectively. Participants’ comfort before EGD was significantly higher in the “4+2” protocol group measured by both the proportion of satisfaction (86.8% vs 63.9%, p=0.002) and the visual analog scale score (median [interquartile range]: 1.0 [1.0–2.0] vs 3.0 [1.0–4.0], p<0.001). The proportion of satisfaction during EGD also significantly improved (59.4% vs 45.4%, p=0.039) in the “4+2” protocol group. The total visibility score was unaffected by the fasting protocol (5.0 [4.0–5.0] vs 4.0 [4.0–5.0], p=0.266). No adverse events were observed during the study. Conclusions: The “4+2” protocol was more comfortable and provided equal mucosal visibility and safety compared with conventional fasting for unsedated EGD. (

Dynamic analysis for gene expression profiles of endothelial colony forming cells under hypoxia

De-Cai Yu,Wen-Du Feng,Xian-Biao Shi,hi-Yong Wang,Wei Ge,Chun-Ping Jiang,Xi-Tai Sun,Yi-Tao Ding 한국유전학회 2013 Genes & Genomics Vol.35 No.4

Previous studies have shown that endothelial colony forming cells (ECFCs) play an important role in the neovascularization of tumors. Hypoxia is emphasized as an important promoter of angiogenesis. However, little is known about genome-wide transcriptional regulation of ECFCs under hypoxic conditions. In this study, gene expression profiles in ECFCswere evaluated under hypoxic conditions for 3, 6, 12, 24,and 48 h, using Affymetrix U133 plus 2.0 chip microarray. 1,103 hypoxia-regulated genes were filtered, with 379(0.693 %) genes up-regulated and 724 (1.32 %) genes downregulated. Most of the up-regulated genes were involved in apoptosis, cell proliferation, or metabolic processes, while most of the down-regulated genes were involved in cell adherence,cell cycle,DNAandmRNAmetabolic processes,multi-cellular organism development, protein metabolic processes, response to stress, signal transduction, or transport. This expression profile is ECFC-specific, because it is significantly different from those of endothelial cells and smooth muscle cells under hypoxic conditions. Moreover, hypoxia-regulated apoptosis in ECFCs is mainly related with the mitochondrial pathway (p53-BAX-Caspase-9) and the death receptor pathway (FASCaspase-8-Caspase-3). MAPK pathway is activated in ECFCs under hypoxic conditions. The differentially expressed genes of ECFCs were identified under hypoxic conditions, and related with cell apoptosis, cell cycle and MAPK pathways, shedding light on the mechanism of angiogenesis.

Triterpenoids and Diarylheptainoids from Alnus hirsuta Inhibit HIF-1 in AGS Cells

Jin, Wen-Yi,Cai, Xing-Fu,Na, Min-Kyun,Lee, Jung-Joon,Bae, Ki-Hwan 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.4

Bioassay-guided fractionation and purification of the EtOAc soluble fraction from the MeOH extract of the stem bark of Alnus hirsuta (Betulaceae), using an in vitro HIF-1 assay, led to the isolation of four triterpenoids (1-4) and six diarylheptanoids (5-10). Their structures were deter-mined by comparison with the physicochemical and spectroscopic data in the literature. These compounds were investigated for their effects on the hypoxia-induced HlF-1 activation using an HIF-1 a mediated reporter gene assay in AGS cells. Among them, two diarylheptanoids, 2-oxatrycyclo[13.2.2.13,7]eicosa-3,5,7(20),15,17,18-hexaen-10-16-diol (6) and 2-oxatrycyclo[13.2.2.13,7]eicosa-3,5,7-(20),15,17,18-hexaen-10-one (7), inhibited HIF-1 activation dose-dependently with IC$_{50}$ values of 11.2 ${\mu}$M and 12.3 ${\mu}$M, respectively. These two compounds had no significant cytotoxicity to the AGS cells at the effective concentration for the inhibition of HIF-1 activation.

RALY RNA Binding Protein-like Reduced Expression is Associated with Poor Prognosis in Clear Cell Renal Cell Carcinoma

Cui, Zhi-Wen,Xia, Ye,Ye, Yi-Wang,Jiang, Zhi-Mao,Wang, Ya-Dong,Wu, Jian-Ting,Sun, Liang,Zhao, Jun,Fa, Ping-Ping,Sun, Xiao-Juan,Gui, Yao-Ting,Cai, Zhi-Ming Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.7

The molecular mechanisms involved in the progression of clear cell renal cell carcinomas (ccRCCs) are still unclear. The aim of this study was to analyse the relationships between expression of RALYL and clinical characteristics. In 41 paired samples of ccRCCs and adjacent normal tissues, we used real-time qPCR to evaluate the expression of RALYL mRNA. RALYL protein levels were determined in 146 samples of ccRCC and 37 adjacent normal tissues by immunohistochemistry. Statistical analysis was used to explore the relationships between expression of RALYL and the clinical characteristics (gender, age, tumor size, T stage, N stage, M stage, survival times and survival outcome) in ccRCC. In addition, these patients were follow-up period 64 months (range: 4~116months) to investigate the influence on prognosis. We found significantly differences between ccRCC tissues and normal tissues (p<0.001, paired-sample t test) in mRNA levels of RALYL. Immunohistochemistry analyses in 146 ccRCC samples and 37 adjacent normal tissues showed significantly lower RALYL protein levels in ccRCC samples (${\chi}^2$-test, p<0.001), inversely correlating with tumour size (p=0.024), T stage (0.005), N stage (p<0.001) as well as M stage (p=0.019), but not age (p=0.357) and gender (p=0.348). Kaplan-Meier survival analysis demonstrated that people with lower level of RALYL expression had a poorer survival rate than those with a higher level of RALYL expression, significantly different by the log-rank test (p=0.011). Cox regression analysis indicated that RALYL expression (p=0.039), N stage (p=0.008) and distant metastasis (p<0.001) were independent prognosis factors for the overall survival of ccRCC patients. We demonstrated that the expression of RALYL was significantly low in ccRCC and correlated with a poor prognosis in a large number of clinical samples. Our findings showed that RALYL may be a potential therapeutic target as well as a poor prognostic factor.

Clinical Implications of p57^KIP2 Expression in Breast Cancer

Xu, Xiao-Yin,Wang, Wen-Qian,Zhang, Lei,Li, Yi-Ming,Tang, Miao,Jiang, Nan,Cai, Shou-Liang,Wei, Liang,Jin, Feng,Chen, Bo Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.10

Objective: To study the relationship between expression of $p57^{KIP2}$ and prognosis and other clinicopathological parameters in invasive breast cancers. Methods: We assessed the expression of $p57^{KIP2}$ in 89 cases of invasive breast cancer and 20 cases of normal breast tissue by immunohistochemical methods and analyzed the results with SPSS software (ver. 16.0). Result: The positive expression rates of $p57^{KIP2}$ protein in the invasive breast cancers and surrounding normal tissue were 30.3% (27/89) and 65% (13/20), respectively. Cases with no $p57^{KIP2}$ expression exhibited a significantly higher post-operative distant metastasis rate than those with $p57^{KIP2}$ expression (37.9% vs. 14.8%; P = 0.01). DFS analysis showed that $p57^{KIP2}$-/C-erbB-2+ tumors also exhibited a significantly higher post-operative distant metastasis rate than the other groups (66.7% vs. 29.2%; P = 0.007), as did $p57^{KIP2}$-/p53+ tumors (64.3% vs. 22.7%; P = 0.001). Survival analysis revealed that $p57^{KIP2}$ was associated with breast cancer-specific survival overall (P = 0.045, log-rank test). Subgroup analysis demonstrated that individuals with $p57^{KIP2}$-/C-erbB-2+tumors experienced significantly worse post-operative survival than those with $p57^{KIP2}$-/C-erbB-2- or other tumors (P = 0.006, log-rank test). $p57^{KIP2}$-/p53+ tumors were associated with significantly worse post-operative survival than $p57^{KIP2}$-/p53- or other tumors (P = 0.001, log-rank test). Cox regression analysis showed that $p57^{KIP2}$ was a non-independent prognostic factor for breast cancer (P = 0.303). Conclusions: $p57^{KIP2}$ is expressed at low levels in invasive breast cancer and is associated with better overall survival rate and disease-free survival in breast cancer patients, but it was a non-independent prognostic factor for breast cancer. Thus, the connection between $p57^{KIP2}$/p53 and $p57^{KIP2}$/C-erbB-2 may provide biomarkers for breast cancer.

Estimation of the load-deformation responses of flanged reinforced concrete shear walls

Bin Wang,Qingxuan Shi,Wen-Zhe Cai,YI-Gong Peng 국제구조공학회 2020 Structural Engineering and Mechanics, An Int'l Jou Vol.73 No.5

As limited well-documented experimental data are available for assessing the attributes of different deformation components of flanged walls, few appropriate models have been established for predicting the inelastic responses of flanged walls, especially those of asymmetrical flanged walls. This study presents the experimental results for three large-scale T-shaped reinforced concrete walls and examines the variations in the flexural, shear, and sliding components of deformation with the total deformation over the entire loading process. Based on the observed deformation behavior, a simple model based on moment-curvature analysis is established to estimate flexural deformations, in which the changes in plastic hinge length are considered and the deformations due to strain penetration are modeled individually. Based on the similar gross shapes of the curvature and shear strain distributions over the wall height, a proportional relationship is established between shear displacement and flexural rotation. By integrating the deformations due to flexure, shear, and strain penetration, a new load-deformation analytical model is proposed for flexure-dominant flanged walls. The proposed model provides engineers with a simple, accurate modeling tool appropriate for routine design work that can be applied to flexural walls with arbitrary sections and is capable of determining displacements at any position over the wall height. By further simplifying the analytical model, a simple procedure for estimating the ultimate displacement capacity of flanged walls is proposed, which will be valuable for performance-based seismic designs and seismic capacity evaluations.

Attenuation of Beryllium Induced Hepatorenal Dysfunction and Oxidative Stress in Rodents by Combined Effect of Gallic Acid and Piperine

Jun-Quan Zhao,Guo-Zhen Du,You-Cai Xiong,Yi-Fu Wen,Monika Bhadauria,Satendra Kumar Nirala 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.12

We determined a minimum effective dose of gallic acid (3,4,5-trihydroxy benzoic acid; 50 mg/ kg, i.p.) and piperine (10 mg/kg, p.o.) through their therapeutic potential and further evaluated them individually and in combination against beryllium-induced biochemical alterations and oxidative stress consequences in female albino rats. The administration of beryllium altered blood biochemical variables by significantly depleting hemoglobin, albumin and urea, whereas it enhanced bilirubin and creatinine. The release of serum transaminase, lactate dehydrogenase and γ-glutamyl transpeptidase was significantly greater, and was concomitant with a decrease in serum alkaline phosphatase. A significant increase in lipid peroxidation and a decrease in glutathione, superoxide dismutase and catalase in the liver and kidney was an indication of oxidative stress due to beryllium exposure. Individual administration of gallic acid and piperine moderately reversed the altered biochemical variables, whereas the combination of these was found to completely reverse the beryllium-induced biochemical alterations and oxidative stress consequences. We concluded that gallic acid exerts a synergistic effect when administered with piperine and provides a more pronounced therapeutic potential in reducing beryllium-induced hepatorenal dysfunction and oxidative stress consequences.