Molecular Cloning of the cDNA of Heat Shock Protein 88 Gene from the Entomopathogenic Fungus, Paecilomyces tenuipes Jocheon-1

( Ya Qi Liu ),( Nam Sook Park ),( Yong Gyun Kim ),( Keun Ki Kim ),( Hyun Chul Park ),( Hong Joo Son ),( Chang Ho Hong ),( Sang Mong Lee ) 한국잠사학회 2014 International Journal of Industrial Entomology Vol.28 No.2

The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomycestenuipes Jocheon -1 was obtained by screening the Paecilomyces tenuipes (P. tenuipes )Jocheon-1 Uni-Zap cDNA library and performing 5` RACE polymerase chain reaction(PCR). The P. tenuipes Jocheon-1 HSP88 cDNA contained an open reading frame (ORF) of2, 139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of theP. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88and 45-76% identity to other fungal homologous HSP88s. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipesJocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade. The P. tenuipes Jocheon-1 HSP88 also contained the conserved ATPase domain at the N-terminalregion. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as an 88 kilodalton(kDa) polypeptide in baculovirus-infected insect Sf9 cells. Under higher temperature conditionsfor the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stressinduced a higher level of mRNA expression compared to normal growth conditions.

Molecular Cloning of the cDNA of Heat Shock Protein 88 Gene from the Entomopathogenic Fungus, Paecilomyces tenuipes Jocheon-1

Liu, Ya-Qi,Park, Nam Sook,Kim, Yong Gyun,Kim, Keun Ki,Park, Hyun Chul,Son, Hong Joo,Hong, Chang Ho,Lee, Sang Mong Korean Society of Sericultural Science 2014 International Journal of Industrial Entomology Vol.28 No.2

The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening the Paecilomyces tenuipes (P. tenuipes) Jocheon-1 Uni-Zap cDNA library and performing 5' RACE polymerase chain reaction (PCR). The P. tenuipes Jocheon-1 HSP88 cDNA contained an open reading frame (ORF) of 2,139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88 and 45-76% identity to other fungal homologous HSP88s. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade. The P. tenuipes Jocheon-1 HSP88 also contained the conserved ATPase domain at the N-terminal region. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as an 88 kilodalton (kDa) polypeptide in baculovirus-infected insect Sf9 cells. Under higher temperature conditions for the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1 HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stress induced a higher level of mRNA expression compared to normal growth conditions.

Comparison of the Genomic Structure of the Heat Shock Protein-88(Hsp88) Genes in the Four Entomopathogenic Fungal Strains, Paecilomyces tenuipes Jocheon-1, P. tenuipes, Cordyceps militaris, and C. pruinosa

( Ya Qi Liu ),( Nam Sook Park ),( Yong Gyun Kim ),( Keun Ki Kim ),( Hyun Chul Park ),( Hong Joo Son ),( Sang Mong Lee ) 한국잠사학회 2012 International Journal of Industrial Entomology Vol.25 No.1

Comparison on the genomic structure and phylogenetic relationship of the Hsp88 genes from P. tenuipes Jochoen-1, P. tenuipes, C. militaris and C. pruinosa was described. The Hsp88 genes from the three entomopathogenic strains, P. tenuipes Jocheon-1(strain), P. tenuipes(original species), and C. militaris contain the identical genomic structure, namely 5 introns and 6 exons with the length of 13, 62, 32, 1,438, 306, 288 nucleotides encoding 713 amino acid residues, whereas in case of C. pruinosa, it contains 4 introns and 5 exons with the length of 13, 62, 32, 1,744, 288 nucleotides encoding 713 amino acid residues. The genomic DNA length of the Hsp88 genes from P. tenuipes Jocheon-1 and P. tenuipes are both 2,600 nucleotides long in size. The Hsp88 genes from C. militaris and C. pruinosa are 2,582, 2,576 nucleotides long in size, respectively. Hsp88 genes of the P. tenuipes Jochoen-1, P. tenuipes, C. militaris and C. pruinosa also contain the conserved ATP-binding domain. Phylogenetic analysis of the Hsp genes of the four strains tested in this study showed that the fungal Hsp88 is divided into two separate clades, ascomycetes and deutromycete. Within the ascomycetes fungal clade, the P. tenuipes Jochoen-1 and P. tenuipes formed a subgroup, on the other hand, C. militaris and C. pruinosa formed another subgroup. Pair-wise comparison of P. tenuipes Jocheon-1 Hsp88 with those of P. tenuipes, C. militaris and C. pruinosa Hsp88s revealed significant identity in deduced amino acid sequence among these strains. The P. tenuipes Jocheon-1 Hsp88 showed 99% identity with the P. tenuipes, 97% identity with the C. militaris, and 98% identity with the C. pruinosa.

Comparison of the Genomic Structure of the Heat Shock Protein-88(Hsp88) Genes in the Four Entomopathogenic Fungal Strains, Paecilomyces tenuipes Jocheon-1, P. tenuipes, Cordyceps militaris, and C. pruinosa

Liu, Ya-Qi,Park, Nam-Sook,Kim, Yong-Gyun,Kim, Keun-Ki,Park, Hyun-Chul,Son, Hong-Joo,Lee, Sang-Mong Korean Society of Sericultural Science 2012 International Journal of Industrial Entomology Vol.25 No.1

Comparison on the genomic structure and phylogenetic relationship of the Hsp88 genes from P. tenuipes Jochoen-1, P. tenuipes, C. militaris and C. pruinosa was described. The Hsp88 genes from the three entomopathogenic strains, P. tenuipes Jocheon-1(strain), P. tenuipes(original species), and C. militaris contain the identical genomic structure, namely 5 introns and 6 exons with the length of 13, 62, 32, 1,438, 306, 288 nucleotides encoding 713 amino acid residues, whereas in case of C. pruinosa, it contains 4 introns and 5 exons with the length of 13, 62, 32, 1,744, 288 nucleotides encoding 713 amino acid residues. The genomic DNA length of the Hsp88 genes from P. tenuipes Jocheon-1 and P. tenuipes are both 2,600 nucleotides long in size. The Hsp88 genes from C. militaris and C. pruinosa are 2,582, 2,576 nucleotides long in size, respectively. Hsp88 genes of the P. tenuipes Jochoen-1, P. tenuipes, C. militaris and C. pruinosa also contain the conserved ATP-binding domain. Phylogenetic analysis of the Hsp genes of the four strains tested in this study showed that the fungal Hsp88 is divided into two separate clades, ascomycetes and deutromycete. Within the ascomycetes fungal clade, the P. tenuipes Jochoen-1 and P. tenuipes formed a subgroup, on the other hand, C. militaris and C. pruinosa formed another subgroup. Pair-wise comparison of P. tenuipes Jocheon-1 Hsp88 with those of P. tenuipes, C. militaris and C. pruinosa Hsp88s revealed significant identity in deduced amino acid sequence among these strains. The P. tenuipes Jocheon-1 Hsp88 showed 99% identity with the P. tenuipes, 97% identity with the C. militaris, and 98% identity with the C. pruinosa.

Matrine Reduces Proliferation of Human Lung Cancer Cells by Inducing Apoptosis and Changing miRNA Expression Profiles

Liu, Yong-Qi,Li, Yi,Qin, Jie,Wang, Qian,She, Ya-Li,Luo, Ya-Li,He, Jian-Xin,Li, Jing-Ya,Xie, Xiao-Dong Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.5

Matrine, a main active component extracted from dry roots of Sophora flavecens, has been reported to exert antitumor effects on A549 human non-small lung cancer cells, but its mechanisms of action remain unclear. To determine effects of matrine on proliferation of A549 cells and assess possible mechanisms, MTT assays were employed to detect cytotoxicity, along with o flow cytometric analysis of DNA content of nuclei of cells following staining with propidium iodide to analyze cell cycle distribution. Western blotting was performed to determined expression levels of Bax, Bcl-2, VEGF and HDAC1, while a microarray was used to assessed changes of miRNA profiles. In the MTT assay, matrine suppressed growth of human lung cancer cell A549 in a dose- and timedependent manner at doses of 0.25-2.5 mg/ml for 24h, 48h or 72h. Matrine induced cell cycle arrest in G0/G1 phase and decreased the G2/M phase, while down-regulating the expression of Bcl2 protein, leading to a reduction in the Bcl-2/Bax ratio. In addition, matrine down regulated the expression level of VEGF and HDAC1 of A549 cells. Microarray analysis demonstrated that matrine altered the expression level of miRNAs compared with untreated control A549 cells. In conclusion, matrine could inhibit proliferation of A549 cells, providing useful information for understanding anticancer mechanisms.

Molecular Cloning, Expression and Characterizaion of Heat Shock Protein Gene from Paecilomyces tenuipes Jocheon-1

Liu Ya Qi,Nam Sook Park,Han Seok Kang,Seon Ku Kim,Yong Gyun Kim,Byung Uuk Cho,Teak Soon Shin,Keun Ki Kim,Hyun Chul Park,Hong Joo Son,Hong Gu Lee,Sang Mong Lee 한국응용곤충학회 2011 한국응용곤충학회 학술대회논문집 Vol.2011 No.05

In this study, a full-length heat shock protein88 complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening of P. tenuipesJocheon-1 Uni-Zap cDNA library and 5' RACE polymerase chain reaction. The Paecilomyces tenuipes Jocheon-1 heat shock protein88 cDNA contains an open reading frame of 2,139 bp encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipes Jocheon-1 HSP88 cDNA showed 77% identity to N. haematococca HSP88 and 45-76% identity to other fungi HSP88. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade and P. tenuipes Jocheon-1 HSP88 also contains the conserved ATPase domain at the N-terminal. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as a 88 kDa polypeptide in baculovirus-infected insect Sf9 cells. Under different stress conditions, mRNA expression of P. tenuipes Jocheon-1 HSP88 were quantified by real-time PCR and the result showed that heat shock stress affected the mRNA expression levels of P. tenuipes Jocheon-1 HSP88.

Genomic Structure of the Heat Shock Protein88 genes of Paecilomyces tenuipes Jocheon-1, Paecilomyces tenuipes, Cordyceps militaris & Cordycepspruinosa

Liu Ya Qi,Nam Sook Park,Han Seok Kang,Seon Ku Kim,Yong Gyun Kim,Byung Uuk Cho,Teak Soon Shin,Keun Ki Kim,Hyun Chul Park,Hong Joo Son,Hong Gu Lee,Sang Mong Lee 한국응용곤충학회 2011 한국응용곤충학회 학술대회논문집 Vol.2011 No.05

The genomic structure and phylogenetic relationships of HSP88 genes from P. tenuipes Jocheon-1, P. tenuipes, C. militaris and C. pruinosa are described. The HSP88 genomic DNA from P. tenuipes Jocheon-1, P. tenuipes and C. militaris all contain 5 introns and 6 exons with the length of 13, 62, 32, 1438, 306, 288 bp, encoding 713 amino acid residues. C. pruinosa HSP88 genomic DNA contains 4 introns and 5 exons encoding 713 amino acids. The length of each exon of C. pruinosa HSP88 is 13, 62, 32, 1744, 288 bp and the length of exon 4 is identical to the total length of exon 4 and exon 5 of HSP88 of P. tenuipes Jochoen-1, P. tenuipes, and C. militaris. The deduced amino acid sequence of P. tenuipes Jocheon-1 HSP88 showed 99% identity with the P. tenuipes, 97% identity with the Cordyceps militaris, and 98% identity with the C. pruinosa. Phylogenetic analysis confirmed that the P. tenuipes Jocheon-1, P. tenuipes, C. militaris and C. pruinosa HSP88 are placed together within the ascomycetes group of fungal clade.

Risk Factors of Unsatisfactory Robot-Assisted Pedicle Screw Placement: A Case-Control Study

Qi Zhang,Ming-Xing Fan,Xiao-Guang Han,Ya-Jun Liu,Da He,Bo Liu,Wei Tian 대한척추신경외과학회 2021 Neurospine Vol.18 No.4

Objective: To identify potential risk factors of unsatisfactory screw position during robot-assisted pedicle screw fixation. Methods: A retrospective analysis of robot-assisted pedicle screw fixation performed in Beijing Jishuitan Hospital from March 2018 to March 2019 was conducted. Research data was collected from the medical record and imaging systems. Univariate tests were performed on the potential risk factors (patient’s characteristics and surgical factors) of unsatisfactory screw position during robot-assisted pedicle screw fixation. For statistically significant variables in univariate tests, a logistic regression test was used to identify independent risk factors for unsatisfactory screw position. Results: A total of 780 pedicle screws placed in 163 robot-assisted surgeries were analyzed. The rate of perfect screw positions was 93.08%, and the unsatisfactory rate was 6.92%. In patients with severe obesity (body mass index≥30 kg/m2) (odds ratio [OR], 2.459; 95% confidence interval [CI], 1.199–5.044; p=0.014), osteoporosis (T≤-2.5) (OR, 1.857; 95% CI, 1.046–3.295; p=0.034), and the segments 3 levels away from the tracker (OR, 2.216; 95% CI, 1.119–4.387; p=0.022), robot-assisted pedicle screw placement has a higher risk of screw malposition. Conclusion: During robot-assisted pedicle screw placement for patients with severe obesity, osteoporosis, and segments 3 levels away from the tracker, vigilance should be maintained during surgery to avoid postoperative complications due to unsatisfactory screw position.

Systemic Analysis on Risk Factors for Breast Cancer Related Lymphedema

Zhu, Ya-Qun,Xie, Yu-Huan,Liu, Feng-Huan,Guo, Qi,Shen, Pei-Pei,Tian, Ye Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.16

Background: To evaluate risk factors for upper extremity lymphedema due to breast cancer surgery. Materials and Methods: Clinical studies published on PubMed, Ovid, EMbase, and Cochrane Library from January 1996 to December 2012 were selected. Results: Twenty-five studies were identified, including 12,104 patients. Six risk factors related to the incidence of lymphedema after breast cancer treatment were detected: axillary lymph node dissection (OR=3.73, 95%CI 1.16 to 11.96), postoperative complications (OR=2.64, 95%CI 1.10 to 6.30), hypertension (OR=1.83, 95%CI 1.38 to 2.42), high body mass index (OR=1.80, 95%CI 1.30 to 2.49), chemotherapy (OR=1.38, 95%CI 1.07 to 1.79) and radiotherapy (OR=1.35, 95%CI 1.10 to 1.66). We found significant protective factors for lymphedema: pathologic T classification (OR=0.57, 95%CI 0.36 to 0.91) and stage (OR=0.60, 95%CI 0.39 to 0.93), while some factors, like age, number of positive lymph nodes, number of lymph node dissection, demonstrated no obvious correlation. Conclusions: Axillary lymph node dissection, postoperative complications, hypertension, body mass index, chemotherapy, radiotherapy are risk factors for lymphedema after breast cancer treatment. Attention should be paid to patients with risk factors to prevent the occurrence of lymphedema.

Application of Crossover Analysis-logistic Regression in the Assessment of Gene- environmental Interactions for Colorectal Cancer

Wu, Ya-Zhou,Yang, Huan,Zhang, Ling,Zhang, Yan-Qi,Liu, Ling,Yi, Dong,Cao, Jia Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.5

Background: Analysis of gene-gene and gene-environment interactions for complex multifactorial human disease faces challenges regarding statistical methodology. One major difficulty is partly due to the limitations of parametric-statistical methods for detection of gene effects that are dependent solely or partially on interactions with other genes or environmental exposures. Based on our previous case-control study in Chongqing of China, we have found increased risk of colorectal cancer exists in individuals carrying a novel homozygous TT at locus rs1329149 and known homozygous AA at locus rs671. Methods: In this study, we proposed statistical method-crossover analysis in combination with logistic regression model, to further analyze our data and focus on assessing gene-environmental interactions for colorectal cancer. Results: The results of the crossover analysis showed that there are possible multiplicative interactions between loci rs671 and rs1329149 with alcohol consumption. Multifactorial logistic regression analysis also validated that loci rs671 and rs1329149 both exhibited a multiplicative interaction with alcohol consumption. Moreover, we also found additive interactions between any pair of two factors (among the four risk factors: gene loci rs671, rs1329149, age and alcohol consumption) through the crossover analysis, which was not evident on logistic regression. Conclusions: In conclusion, the method based on crossover analysis-logistic regression is successful in assessing additive and multiplicative gene-environment interactions, and in revealing synergistic effects of gene loci rs671 and rs1329149 with alcohol consumption in the pathogenesis and development of colorectal cancer.