Attenuated Secretion of the Thermostable Xylanase xynB from Pichia pastoris Using Synthesized Sequences Optimized from the Preferred Codon Usage in Yeast

( Huang Yuan Kai ),( Yao Sheng Chen ),( De Lin Mo ),( Pei Qing Cong ),( Zu Yong He ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.3

Xylanase has been used extensively in the industrial and agricultural fields. However, the low-yield production of xylanase from native species cannot meet the increasing demand of the market. Therefore, improving the heterologous expression of xylanase through basic gene optimization may help to overcome the shortage. In this study, we synthesized a high-GC-content native sequence of the thermostable xylanase gene xynB from Streptomyces olivaceoviridis A1 and, also designed a slightly AT-biased sequence with codons completely optimized to be favorable to Pichia pastoris. The comparison of the sequences` expression efficiencies in P. pastoris X33 was determined through the detection of single-copy-number integrants, which were quantified using qPCR. Surprisingly, the high GC content did not appear to be detrimental to the heterologous expression of xynB in yeast, whereas the optimized sequence, with its extremely skewed codon usage, exhibited more abundant accumulation of synthesized recombinant proteins in the yeast cell, but an approximately 30% reduction of the secretion level, deduced from the enzymatic activity assay. In this study, we developed a more accurate method for comparing the expression levels of individual yeast transformants. Moreover, our results provide a practical example for further investigation of what constitutes a rational design strategy for a heterologously expressed and secreted protein.

Integrin-linked Kinase Functions as a Tumor Promoter in Bladder Transitional Cell Carcinoma

Wang, De-Lin,Lan, Jian-Hua,Chen, Liang,Huang, Biao,Li, Zeng,Zhao, Xiu-Min,Ma, Qiang,Sheng, Xia,Li, Wen-Bin,Tang, Wei-Xue Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.6

The aim of this study was to elucidate the role of the integrin-linked kinase (ILK) gene in development of human bladder transitional cell carcinoma (BTCC). Expression of ILK protein and ILK mRNA in 56 cases of human BTCC tissue and in 30 cases of adjacent normal bladder tissue was detected by immunohistochemistry S-P and reverse transcription polymerase chain reaction (RT-PCR), respectively. Four specific miRNA RNAi vectors targeting human ILK were synthesized and transfected into BIU-87 cells by liposome to obtain stable expression cell strains. The influence of ILK on proliferation of BTCC was detected by MTT, FCM on athymic mouse tumorigenesis. The positive rate of ILK protein in BTCC tissue (53.6%) was much higher than adjacent normal bladder tissue (10.0%) (p<0.05). Similarly, expression of ILK mRNA in BTCC tissue ($0.540{\pm}0.083$) was significantly higher than in adjacent normal bladder tissue ($0.492{\pm}0.070$) (p<0.05). MTT showed that the proliferation ability of miRNA-ILK transfected group was clearly decreased (p<0.05), the cell cycle being arrested in G0/G1-S, an tumorigenesis in vivo was also significantly reduced (p<0.05). ILK gene transcription and protein expression may be involved in the development of BTCC, so that ILK might be the new marker for early diagnosis and the new target for gene treatment.

Introducer Curving Technique for the Prevention of Tilting of Transfemoral Günther Tulip Inferior Vena Cava Filter

Liang Xiao,De-sheng Huang,Jing Shen,Jia-jie Tong 대한영상의학회 2012 Korean Journal of Radiology Vol.13 No.4

Objective: To determine whether the introducer curving technique is useful in decreasing the degree of tilting of transfemoral Tulip filters. Materials and Methods: The study sample group consisted of 108 patients with deep vein thrombosis who were enrolled and planned to undergo thrombolysis, and who accepted transfemoral Tulip filter insertion procedure. The patients were randomly divided into Group C and Group T. The introducer curving technique was Adopted in Group T. The post-implantation filter tilting angle (ACF) was measured in an anteroposterior projection. The retrieval hook adhering to the vascular wall was measured via tangential cavogram during retrieval. Results: The overall average ACF was 5.8 ± 4.14 degrees. In Group C, the average ACF was 7.1 ± 4.52 degrees. In Group T, the average ACF was 4.4 ± 3.20 degrees. The groups displayed a statistically significant difference (t = 3.573, p = 0.001) in ACF. Additionally, the difference of ACF between the left and right approaches turned out to be statistically significant (7.1 ± 4.59 vs. 5.1 ± 3.82, t = 2.301, p = 0.023). The proportion of severe tilt (ACF ≥ 10º) in Group T was significantly lower than that in Group C (9.3% vs. 24.1%, χ2 = 4.267, p = 0.039). Between the groups, the difference in the rate of the retrieval hook adhering to the vascular wall was also statistically significant (2.9% vs. 24.2%, χ2 = 5.030, p = 0.025). Conclusion: The introducer curving technique appears to minimize the incidence and extent of transfemoral Tulip filter tilting. Objective: To determine whether the introducer curving technique is useful in decreasing the degree of tilting of transfemoral Tulip filters. Materials and Methods: The study sample group consisted of 108 patients with deep vein thrombosis who were enrolled and planned to undergo thrombolysis, and who accepted transfemoral Tulip filter insertion procedure. The patients were randomly divided into Group C and Group T. The introducer curving technique was Adopted in Group T. The post-implantation filter tilting angle (ACF) was measured in an anteroposterior projection. The retrieval hook adhering to the vascular wall was measured via tangential cavogram during retrieval. Results: The overall average ACF was 5.8 ± 4.14 degrees. In Group C, the average ACF was 7.1 ± 4.52 degrees. In Group T, the average ACF was 4.4 ± 3.20 degrees. The groups displayed a statistically significant difference (t = 3.573, p = 0.001) in ACF. Additionally, the difference of ACF between the left and right approaches turned out to be statistically significant (7.1 ± 4.59 vs. 5.1 ± 3.82, t = 2.301, p = 0.023). The proportion of severe tilt (ACF ≥ 10º) in Group T was significantly lower than that in Group C (9.3% vs. 24.1%, χ2 = 4.267, p = 0.039). Between the groups, the difference in the rate of the retrieval hook adhering to the vascular wall was also statistically significant (2.9% vs. 24.2%, χ2 = 5.030, p = 0.025). Conclusion: The introducer curving technique appears to minimize the incidence and extent of transfemoral Tulip filter tilting.

Prediction of Length of ICU Stay Using Data-mining Techniques: an Example of Old Critically Ill Postoperative Gastric Cancer Patients

Zhang, Xiao-Chun,Zhang, Zhi-Dan,Huang, De-Sheng Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.1

Objective: With the background of aging population in China and advances in clinical medicine, the amount of operations on old patients increases correspondingly, which imposes increasing challenges to critical care medicine and geriatrics. The study was designed to describe information on the length of ICU stay from a single institution experience of old critically ill gastric cancer patients after surgery and the framework of incorporating data-mining techniques into the prediction. Methods: A retrospective design was adopted to collect the consecutive data about patients aged 60 or over with a gastric cancer diagnosis after surgery in an adult intensive care unit in a medical university hospital in Shenyang, China, from January 2010 to March 2011. Characteristics of patients and the length their ICU stay were gathered for analysis by univariate and multivariate Cox regression to examine the relationship with potential candidate factors. A regression tree was constructed to predict the length of ICU stay and explore the important indicators. Results: Multivariate Cox analysis found that shock and nutrition support need were statistically significant risk factors for prolonged length of ICU stay. Altogether, eight variables entered the regression model, including age, APACHE II score, SOFA score, shock, respiratory system dysfunction, circulation system dysfunction, diabetes and nutrition support need. The regression tree indicated comorbidity of two or more kinds of shock as the most important factor for prolonged length of ICU stay in the studied sample. Conclusions: Comorbidity of two or more kinds of shock is the most important factor of length of ICU stay in the studied sample. Since there are differences of ICU patient characteristics between wards and hospitals, consideration of the data-mining technique should be given by the intensivists as a length of ICU stay prediction tool.

MLH1 Polymorphisms and Cancer risk: a Meta-analysis Based on 33 Case-control Studies

Xu, Jia-Li,Yin, Zhi-Qiang,Huang, Ming-De,Wang, Xie-Feng,Gao, Wen,Liu, Ling-Xiang,Wang, Rong-Sheng,Huang, Pu-Wen,Yin, Yong-Mei,Liu, Ping,Shu, Yong-Qian Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.3

Objective: Cumulative evidence suggests that MLH1, the key component in the mismatch pathway, plays an important role in human cancers. Two potential functional polymorphisms (-93G>A and I219V) of MLH1 have been implicated in cancer risk. The aim of this meta-analysis was to summarize the evidence for associations. Methods: Eligible studies were identified by searching the electronic literature PubMed, ScienceDirect and Embase databases for relevant reports and bibliographies. Studies were included if of case-control design investigating MLH1 polymorphisms (-93G>A and I219V) and cancer risk with sufficient raw data for analysis. Odds ratios (OR) and 95% confidence intervals (95% CI) were used to evaluate the strength of associations. Results: Our meta-analysis from 33 published case-control studies showed the variant A allele of -93G>A polymorphism to be associated with increased risk in all genetic models (AA vs. GG: OR = 1.22, 95% CI: 1.03-1.44), especially among non-Asians (AA vs. GG: OR = 1.28, 95% CI: 1.04-1.58). For the I219V polymorphism, however, there was no main effect associated with overall cancer risk in any genetic model. Conclusions: The meta-analysis suggested that the MLH1 -93G>A polymorphism may be a biomarker of cancer susceptibility. Large sample association studies and assessment of gene-to-gene as well as gene-to-environment interactions are required to confirm these findings.

Baicalin attenuates adriamycin-induced nephrotic syndrome by regulating fibrosis procession and inflammatory reaction

Tan Ning,Sun Chen-Xia,Zhu Hui-Jun,Li De-Yu,Huang Sheng-Guang,He Shou-Di 한국유전학회 2021 Genes & Genomics Vol.43 No.9

Background Baicalin has anti-infammatory, antibacterial, blood platelet aggregation-inhibiting, free oxygen radical-clearing, and endotoxin-decreasing properties. However, its molecular mechanism involved in the treatment of Adriamycin-induced nephrotic syndrome (NS) is still unclear. Objective This study aimed to explore the efects of baicalin on Adriamycin-induced nephrotic syndrome (NS) and to characterize the genes involved in this progression. Methods We established Adriamycin-induced NS model in 32 rats and used six rats in Sham group. Urinary total protein content and creatinine serum were assessed as physiological indicators. H&E staining was used to observe the pathological changes. We determined gene expression profles using transcriptome sequencing in the rat kidney tissues from Sham, Adriamycin, and Adriamycin+baicalin groups. KEGG was carried out to analyze the enriched pathways of diferentially expressed genes among these groups. Results Baicalin treatment relieved renal injury in NS rats. Expression of 363 genes was signifcantly diferent between the Adriamycin and Adriamycin+baicalin M groups. Most of the diferentially expressed genes were enriched in pathways involved in epithelial-mesenchymal transition (EMT), fbrosis, apoptosis, and infammation. Conclusions Overall, these data suggest that Adriamycin-induced NS can be attenuated by baicalin through the suppression of fbrosis-related genes and infammatory reactions. Baicalin is a potential drug candidate for the treatment of NS, and the identifed genes represent potential therapeutic targets.

Association between the TP53BP1 rs2602141 A/C Polymorphism and Cancer Risk: A Systematic Review and Meta-Analysis

Liu, Lei,Zhang, Dong,Jiao, Jing-Hua,Wang, Yu,Wu, Jing-Yang,Huang, De-Sheng Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

Background: The p53-binding protein 1 (TP53BP1) gene may be involved in the development of cancer through disrupting DNA repair. However, investigation of associations between TP53BP1 rs2602141 A/C polymorphism and cancer have yielded contradictory and inconclusive outcomes. We therefore performed a meta-analysis to evaluate the association between the TP53BP1 rs2602141 A/C polymorphism and cancer susceptibility. Materials and Methods: Published literature from PubMed, Medline, the Cochrane Library, EMbase, Web of Science, Google (scholar), CBMDisc, Chongqing VIP database, and CNKI database were retrieved. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated using fixed or random-effects models. Publication bias was estimated using funnel plots, Begg's and Egger's test. Results: A total of seven studies (3,018 cases and 5,548 controls) were included in the meta-analysis. Our results showed that the genotype distribution of TP53BP1 rs2602141 A/C was not associated with cancer risk overall. However, on subgroup analysis, we found that TP53BP1 rs2602141 A/C was associated with cancer risk within an allele model (A vs C, OR=1.14, 95%CI: 1.01-1.29) and a codominant model (AA vs CC, OR=1.36, 95%CI: 1.06-1.74) in Asians rather than in Caucasians. Subgroup analysis by cancer type, genotype, and with or without adjustment for controls showed no significant association. Conclusions: The findings suggested an association between rs2602141 A/C polymorphism in TP53BP1 gene and increased risk of cancer in Asians.