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BmNPV chitinase gene deletion enhances foreign gene expression in a BmN cell system
Fang WANG,Chuan-Xi ZHANG 한국곤충학회 2009 Entomological Research Vol.39 No.1
Bombyx mori nucleopolyhedrovirus (BmNPV) is extensively being studied as an expression vector for heterologous gene expression in silkworm-derived cells as well as in the host larvae or pupae. BmNPV chitinase is necessary for liquefaction of the virus-infected host insect. The influence of chitinase on the efficiency of foreign gene expression was studied to provide a scientific basis for improving the BmNPV expression system. The BmNPV chitinase gene (chiA) was deleted and the expression level of the polyhedrin promoter controlling the lacZ gene in BmN cells was determined. Sodium dodecylsulfate (SDS)–olyacrylamide gel electrophoresis (PAGE) showed that β-galactosidase accounted for approximately 6.9 and 7.7% of the total protein in BmN cells infected with the chiA deficient Bm lacZ+chiA−,at 3 and 4 days post infection, while the total protein was 3.2 and 4.2% in cells infected with Bm lacZ+. The relative β-galactosidase activities in Bm lacZ+chiA−[기호] -infected cells improved 2.33- and 1.56-fold compared to those of Bm lacZ+-infected cells at 3 and 4 days post infection. The results of the present study suggest that chitinase deletion could improve the lacZ expression level in the BmNPV-BmN cell expression system.
Wang, Xiao,Song, Zheng-Fang,Xie, Rui-Meng,Pei, Jiao,Xiang, Ming-Fei,Wang, Huan Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.7
Objectives: To analyze underlying disease, fatality rate and the major causes of death of in-patients with malignant tumors in Sichuan Cancer Hospital. Methods: Clinical data of in-patients from 2002 to 2012 were retrospectively analyzed. Results: The top 10 tumors (82.0%of the total) of the malignant tumors of the in-patients were lung, cervical, esophagus, breast, colorectal, nasopharynx, liver and gastric cancers, lymphomas and ovarian cancers. The overall fatality rate was 2.7% during these eleven years, 3.4% and 2.0% for male and females, respectively with statistical significance for the difference (${\chi}^2$=164.737, P<0.001). The top 10 death causes were lung cancer, liver cancer, colorectal cancer, esophagus cancer, gastric cancer, lymphoma, breast cancer, pancreatic cancer, ovarian cancer and nasopharynx cancer. In-patients with pancreatic cancer had the highest fatality rate (9.6%). There were different ranks of death causes in different sex groups and age groups. Conclusion: Prevention and control work of cancer should be enhanced not only for cancers with high incidence such as lung cancer, esophageal cancer but also for the cancers which have low incidence but high fatality rate, such as pancreatic cancer and gallbladder cancer, which would help to improve the survival rate and quality of life of cancer patients in the future.
Expression and Effects of JMJD2A Histone Demethylase in Endometrial Carcinoma
Wang, Hong-Li,Liu, Mei-Mei,Ma, Xin,Fang, Lei,Zhang, Zong-Feng,Song, Tie-Fang,Gao, Jia-Yin,Kuang, Ye,Jiang, Jing,Li, Lin,Wang, Yang-Yang,Li, Pei-Ling Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.7
Previous studies have demonstrated that JMJD2A is a potential oncogene and is overexpressed in human tumors. However, its role in the endometrial carcinoma remains largely unknown. In this study, we discovered that JMJD2A was overexpressed in endometrial carcinoma, using immunohistochemistry, quantitative realtime polymerase chain reaction, and western blotting. Downregulation of JMJD2A led to reduced endometrial carcinoma RL95-2 and ISK cell proliferation, invasion and metastasis as asessed with cell counting kit-8, cell migration and invasive assays. Collectively, our results support that JMJD2A is a promoter of endometrial carcinoma cell proliferation and survival, and is a potential novel drug target.
Wang, Hang-Hui,Song, Yi-Xin,Bai, Min,Jin, Li-Fang,Gu, Ji-Ying,Su, Yi-Jin,Liu, Long,Jia, Chao,Du, Lian-Fang Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.3
The aim was to determine whether ultrasound targeted microbubble destruction (UTMD) promotes dual targeting of HSP72 and HSC70 for therapy of castration-resistant prostate cancer (CRPC), to improve the specific and efficient delivery of siRNA, to induce tumor cell specific apoptosis, and to find new therapeutic targets specific of CRPC.VCaP cells were transfected with siRNA oligonucleotides. HSP70, HSP90 and cleaved caspase-3 expression were determined by real-time quantitative polymerase chain reaction and Western blotting. Apoptosis and transfection efficiency were assessed by flow cytometry. Cell viability assays were used to evaluate safety. We found HSP72, HSC70 and HSP90 expression to be absent or weak in normal prostate epithelial cells (RWPE-1), but uniformly strong in prostate cancerous cells (VCaP). UTMD combined with dual targeting of HSP72 and HSC70 siRNA improve the efficiency of transfection, cell uptake of siRNA, downregulation of HSP70 and HSP90 expression in VCaP cells at the mRNA and protein level, and induction of extensive tumor-specific apoptosis. Cell counting kit-8 assays showed decreased cellular viability in the HSP72/HSC70-siRNA silenced group. These results suggest that the combination of UTMD with dual targeting HSP70 therapy for PCa may be most efficacious, providng a novel, reliable, non-invasive, safe targeted approach to improve the specific and efficient delivery of siRNA, and achieve maximal effects.
Gut Microbiota Community and Its Assembly Associated with Age and Diet in Chinese Centenarians
( Fang Wang ),( Ting Yu ),( Guo Hong Huang ),( Da Cai ),( Xiaolin Liang ),( Hai Yan Su ),( Zhenjun Zhu ),( Danlei Li ),( Yang Yang ),( Pei Hong Shen ),( Rui Feng Mao ),( Lian Yu ),( Mou Ming Zhao ),( 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.8
Increasing evidence suggests that gut microbiota underpin the development of health and longevity. However, our understanding of what influences the composition of this community of the longevous has not been adequately described. Therefore, illumina sequencing analysis was performed on the gut microbiota of centenarians (aged 100-108 years; RC) and younger elderlies (aged 85-99 years; RE) living in Bama County, Guangxi, China and the elderlies (aged 80-92 years; CE) living in Nanning City, Guangxi, China. In addition, their diet was monitored using a semiquantitative dietary questionary (FFQ 23). The results revealed the abundance of Roseburia and Escherichia was significantly greater, whereas that of Lactobacillus, Faecalibacterium, Parabacteroides, Butyricimonas, Coprococcus, Megamonas, Mitsuokella, Sutterella, and Akkermansia was significantly less in centenarians at the genus level. Both clustering analysis and UniFraq distance analysis showed structural segregation with age and diet among the three populations. Using partial least square discriminate analysis and redundancy analysis, we identified 33 and 34 operational taxonomic units (OTUs) as key OTUs that were significantly associated with age and diet, respectively. Age-related OTUs were characterized as Ruminococcaceae, Clostridiaceae, and Lachnospiraceae, and the former two were increased in the centenarians; diet-related OTUs were classified as Bacteroidales, Lachnospiraceae, and Ruminococcaceae. The former two were deceased, whereas the later one was increased, in the high-fiber diet. The age and high-fiber diet were concomitant with changes in the gut microbiota of centenarians, suggesting that age and high-fiber diet can establish a new structurally balanced architecture of gut microbiota that may benefit the health of centenarians.
Fang, Minfeng,Wang, Hui,Wu, Yang,Wang, Qilin,Zhao, Xinfeng,Zheng, Xiaohui,Wang, Shixiang,Zhao, Guifang Korean Chemical Society 2013 Bulletin of the Korean Chemical Society Vol.34 No.6
A rapid and sensitive method for determining usnic acid of Lethariella cladonioides in rat was established using high performance liquid chromatography (HPLC) quadrupole time-of-flight (QTOF) tandem mass (MS/MS). Rat plasma was pretreated by mixture of acetonitrile and chloroform to precipitate plasma proteins. Chromatographic separation was achieved on a column ($50{\times}2.1$ mm, $5{\mu}m$) with a mobile phase consisting of water (containing $5{\times}10^{-3}$ M ammonium formate, pH was adjusted to 3.0 with formic acid) and acetonitrile (20:80, v/v) at a flow rate of 0.3 mL/min. A tandem mass spectrometric detection with an electrospray ionization (ESI) interface was conducted via collision induced dissociation (CID) under negative ionization mode. The MS/MS transitions monitored were m/z 343.0448 ${\rightarrow}$ m/z 313.2017 for usnic acid and m/z 153.1024 ${\rightarrow}$ m/z 136.2136 for protocatechuic acid (internal standard). The linear range was calculated to be 2.0-160.0 ng/mL with a detection limit of 3.0 pg/mL. The inter- and intra-day accuracy and precision were within ${\pm}7.0%$. Pharmacokinetic study showed that the apartment of usnic acid in vivo confirmed to be a two compartment open model. The method was fully valid and will probably be an alternative for pharmacokinetic study of usnic acid.
Wang, Li-Ying,Wang, Xiu-Hua,Tan, Jia-Lian,Xia, Shuai,Sun, Heng-Zhi,Shi, Jin-Wen,Jiang, Ming-Dong,Fang, Liang,Zuo, Hua,Dupati, Gautam,Jang, Kiwan,Shin, Dong-Soo Korean Chemical Society 2012 Bulletin of the Korean Chemical Society Vol.33 No.11
A number of novel small molecules, safrole oxide derivatives 4a-c, 6a-c, 9a-h, were synthesized by the reaction of safrole oxide with anilines 3 and 5, or its alkyl allyl ether derivative 7 with alkyl bromide 8 in moderate yields. The antiproliferative effects of all the target molecules on A549 cell growth were investigated and it was found that the 14 novel compounds could suppress A549 lung cancer cell growth. Among them, compound 6b was the most effective compound in inhibiting the proliferation of A549 cells.